TW200826981A - Tissue plasminogen activator variant uses - Google Patents

Abstract

A method is disclosed for using tenecteplase to restore function in dysfunctional hemodialysis catheters, which have a blood flow rate of less than 300 mL/minute. Kits are also provided with instructions to direct the user to administer tenecteplase in a total dose of about 3 to 4 mg locally into all catheter lumens and allow the tenecteplase to dwell in the catheter for from about one hour to about 72 hours.

Description

200826981 IX. INSTRUCTIONS OF THE INVENTION: TECHNICAL FIELD OF THE INVENTION The present invention relates to the use of tissue plasminogen activator variants which are used to restore the function of a functionally abnormal hemodialysis catheter. [Prior Art] The plasminogen activator is a peptide bond between the cleavage plasmin amino acid residues 561 and 562, which is converted into a plasmin enzyme. Cellulolytic enzyme is an active serine protease that degrades a variety of proteins including fibrin. Currently, the United States has approved five plasminogen activators for the treatment of coronary arterial thrombosis, but none of them have been approved by the FDA for catheter-guided thrombolysis. In the past three years, a large number of clinical studies have been conducted on catheter-guided therapy (CDT) using recombinantly obtained agents. Techniques have been improved and it has been reported that treatment of deep vein thrombosis (DVT) with all available plasminogen activators is effective and safe in non-randomized, uncontrolled observational studies (Elsharawy and Elzayat, Eur J, Vase. Endovasc. Surg·, 24: 209-214 (2002); Semba Anti-Dake, Radiology, 191: 487-494 (1994); Chang et al., /· Vase. and “Mountain····, 12: 247-252 (2001 Castaneda et al., J, Vase. Interv. Radiol., 13: 577-580 (2002) i Semba et al., Tec/z. Fasc. hierv., 4: 99-106 (2001); Allie et al. Dm. J. Card/o/., 90 (suppl 6A): 108H (2002). In general, for an overview of third-generation thrombolytic drugs, see Verstraete, dm. J. MW·, 109: 52-58 (2000). A review of early literature showed that the main complication rate of thrombolysis of peripheral arterial occlusive disease with recombinant tissue plasminogen activator (tPA) was 126382.doc 200826981 5.1% (Semba et al., "/· Vase. Interv. Radiol,, 11: 149-161 (2000); Swischuk et al., /./werv. 12: 423-430 (2001)). tPA test at 0.04 mg/kg/hr dose found 13 % of the Lord Complications (Arepally et al., /·Factory “%·/Werv. 13: 45-50 (2002)) The initial results of reteplase in the treatment of acute lower extremity arterial occlusion show the current use of 0.5 u/ The hr low-dose regimen has a mortality rate of 6% (Davidian et al., 丄Vase. Interv. Radiol., 11: 289-294 (2000)). In recent years, a preliminary study of reteplase using thrombolytics for deep vein thrombosis has reported a major complication rate of 4% (the aforementioned Castaneda fact). Tenecteplase (TNK, TNKASETM, Genentech, Inc., South San Francisco, CA) is a tissue plasminogen activator that complements the modification of native human tissue plasminogen activator A sterile purified glycoprotein of 527 amino acids produced by DNA. These modifications result in molecules having amino acid substitutions at three sites: aspartic acid substituted threonate 103, nopaline acid substituted aspartic acid 117 and four alanine substituted 296-299 amine groups Acid (ionic acid, histidine, arginine and arginine). Tenecteplase converts plasminogen to fibrinolytic enzyme in the presence of fibrin, and the conversion of plasmin to plasmin is limited in the absence of fibrin. Tenepase binds to fibrin in the thrombus and converts plasminogen to cellulolytic enzyme. This initiates localized proteolysis of fibrin associated with blood thrombus, while systemic fibrinogen protein hydrolysis is limited. Tenectease has the same mechanism of action as alteplase and has been shown to potently and effectively promote the in vivo coagulation lysis of 126382.doc 200826981 (Refino et al, Thromb Haemost, 69(6): 841 (1993); Keyt et al., corp. Roc Natl Acad Sci. USA 91:3670-4 (1994) ) In preclinical studies, tenecteplase has demonstrated increased potency and higher fibrin specificity compared with alteplase. Sexual, resistant to plasminogen activator inhibitor (PAI-1) and faster coagulation, while systemic fibrinolysis, plasminogen, and hemorrhage are lower (Refino et al., Thromb· 70: 3 13_3 1 9 (1993); above Keyt et al;

Collen et al., 72: 98-104 (1994); Patel et al. Fasc·/wierv. 12: 559-570 (2001);

Benedict et al., C/rcw/α" (10), 92: 3032-3040 (1995)). In human clinical trials for the treatment of acute myocardial infarction (ami), tenecteplase demonstrated similar efficacy to alteplase, but with a 22% reduction in blood loss, a 23% reduction in blood transfusion and a 16% reduction in secondary bleeding (new Assessment of the Safety and Efficacy of a New Thrombolytic Investigators (ASSENT-2) o Single-bolus tenecteplase compared with front-loaded alteplase in acute myocardial infarction: the ASSENT-2 double blind randomized Trial. Lancet, 354: 716-722 (1999)). There was no significant difference in the rate of intracranial hemorrhage (0.9%). Subjects with a 6-hour symptomatic onset of AMI are eligible for this study. The primary objective was to compare the mortality of subjects 30 days after treatment. Safety endpoints include stroke, hospitalized myocardial reinfarction or pulmonary edema/cardiogenic shock, intracranial hemorrhage (ICH), major bleeding (defined as bleeding requiring hemodynamics or impaired hemodynamics), and severity 126382.doc 200826981 event The ratio. There was no difference in the 30-day mortality between tenecteplase and alteplase in the group of eight patients with eight milk counts evaluated. In addition, there was no difference in the rate between the subjects treated with tenepase and altinase ((93%敎94%, respectively). However, there were two significant non-cerebral lambda hemorrhage events in subjects treated with tenectease compared with those treated with atripizyme (4.66% versus 5 94%, respectively; ρ = 〇〇〇 〇 2) and less blood transfusion (4.25% vs. 5.493⁄4; ρ=〇·〇〇〇2, respectively). <1〇/〇 过敏 Allergic reactions (such as allergic reactions, angioedema, laryngeal edema, rash, and urticaria) were reported in subjects treated with tenecteplase. <〇1% of subjects treated with nepeptidase reported an allergic reaction; however, the cause was not determined. As a result of this study, it is currently approved that tenapopeptidase administered in a weighted dose ranging from 3 to 5 mg in a single intravenous bolus injection is used to reduce mortality associated with acute myocardial infarction (AMI). Due to the excellent safety profile seen in ami and its increased coagulation solubility, researchers have explored the use of tenecteplase as a substitute for thrombolytic agents in non-coronary applications (Semba et al.; Tec/2· Fasc, supra) / price erv. (2001); Sze et al, J. 12: 1456-1457 (2001); Razavi et al, /· Fasc· /Werv., 13: (2), Part 2: S11 (Feb. 2002 ); Nehme et al., J. TVzierv· 7? Kou Shan (9/·, 13: S109 (2002)) 〇

Allie et al. Tenecteplase in Peripheral Thrombolysis: Initial Safety and Feasibility Experience, Society of Interventional Radiology Summary 48, March 2003 (page S17) Rev. 126382.doc 200826981 Ding Lian, Λ 不 普 酶 输 〇 〇 〇 〇 5 〇 mg / hr) is a safe and feasible treatment for peripheral chemical thrombolysis. In addition, it was found that tenectease diluted to 0.0125 mg/m 丨 solution is a viable treatment for occlusion of peripheral arteries and venous thrombosis, which has only a modest effect on fibrinogen content (kart et al., j (3) ι〇99 Ιι〇2

c; ()) And the combination of eptifibatide (eptifibatide) found to be a feasible treatment for acute peripheral stenosis and venous thrombosis ((4) such as et al, Yi/... (4) and cockroach /, 14: 729 733 (2〇〇3)) 〇

Nehme# In, j η% As Hua ~ l3: si〇9 (2〇〇2) provided, estimated that Neptidase makes 14 of the 21 patients with thrombosis of the arteriovenous Teflon HD graft Preliminary results in the study of the efficacy of coagulation. Using a dissolution and waiting technique, 2 telectinase and 3 〇〇〇 U heparin were injected into the graft via blood vessels (4). The duration of drug treatment was not disclosed, but the authors stated that the average private sequence time was 65 minutes. The technical success rate defined as complete graft re-pass was 95% (2 in 21 grafts) and was defined as the clinical success rate of a successful HD after treatment of 9〇% (19 out of 21). Pulse recovery (6 of 21) in 28% of grafts before other mechanical thrombosis. The authors reported a secondary bleeding event at the previous graft site.丄为··C0//· Ca心·〇/,46: 793_8 (2〇〇5) evaluated 85 pre-existing patients with chronic total obstruction>3 months and percutaneous coronary intervention (PCI) The safety and efficacy of intracoronary thrombolysis in unsuccessful subjects was attempted. The subject received a weight-adjusted dose (2_5 mg/hr) of alteplase (n=61) or a standard dose (〇5 mg/hr) of tenep. 126382.doc 200826981 enzyme (n=24) up to 8 Hours, followed by PCI; total dose was dispensed between the guiding catheter and the 3-French coronary infusion catheter. After intracoronary thrombolysis, repetitive PCI was achieved in 54% of all subjects (two treatment groups combined). By multivariate analysis, lesion tapering and lack of bridging collateral were the only successful predictors. Adverse events included hematoma (8% of all subjects) and bleeding requiring blood transfusion (3.5% of all subjects). Tenectease is commercially available in a 50 mg vial and is approved for single bolus administration in patients with AMI (TNKASETM. Complete I square arm 2002 Physicians Desk Reference) , Thomas Medical Economics Co., Montvale, NJ.). When used for approved indications, tenecteplase was reconstituted with sterile water to obtain a final concentration of 5 mg/mL and administered intravenously in a single weight-adjusted bolus injection. As assessed by the ability to draw blood, CATHFLC^ACTIVASE® (alteplase) is indicated for use in restoring the function of a central venous access (CVA) device. The approval is based on two key Genentech-initiated clinical trials of alteplase, which is used to restore catheter function in adults and children over 2 years of age. Subsequently, a third trial of a child subject (age & 17 years old, including certain ages < 2 years old) was performed. All three studies, whether placebo-controlled, double-blind or open-labeled, demonstrated that alteplase was administered at up to 2 mg/2 mL for up to two doses (up to 120 minutes after each administration) Safe and effective treatment for catheter function in adult and pediatric patients with obstructed CVA catheters. After administration of the first dose of alteripine 126382.doc -10- 200826981 enzyme, the recovery rate of the function of the dysfunctional catheter after a retention time of up to 120 minutes was 73.9°/〇-76.5% for the 22-year-old subject and for <6 years old subject was 69.1%. The recovery rate of function after administration of up to two doses of alteplase was 83.5%-89.9% for 22-year-old subjects and 80.0% for <2 years old subjects. A total of 39 out of 1,454 subjects during the three study periods reported serious adverse events. All but three of the severe adverse events were judged to be unrelated to alteplase. No ICH, thrombotic events, or alteplase-related major bleeding were reported during the trial. The most common serious adverse event in these trials was sepsis/bacteria (18%). For peripheral catheter-guided thrombolytic therapy, lyophilized tenecteplase (5 mg/mL) was reconstituted with sterile water and further diluted with standard physiological saline (0.01 to 0.25 mg/mL) (Semba et al., supra, Fa%·/Wav. Radiol·, (2001); Allie et al., J. Cardzo/; above Razavi et al; Semba et al., “/· Fasc·山·〇/·, 13: (2), Part II: S75 (February 2002)). In particular, Razavi et al. previously reported the use of a 0.1 mg/mL dilution of tepeptidase at 25 to 50 mL/hr (0.25-0.5 mg/hr) in standard physiological saline in arteries and veins. Angiographic efficacy is produced in coagulation dissolution (Razavi et al., supra). In addition, Razavi et al, J. Ther., 9:593-598 (2002) disclose that such doses of tenecteplase are safe and effective in peripheral catheter-guided thrombolytic therapy for arterial occlusion and deep vein thrombosis. According to the United States Renal Data System, more than 440,000 people in the United States had end stage renal disease at the end of 2003 (National Institutes of Health, US Kidney Information 126382.doc -11 - 200826981 System USRDS 2005 Data report: American advanced kidney disease episode. Bethesda (MD): National Institute of Diabetes and Digestive and Kidney Diseases (2005). Most of these patients undergo regular hemodialysis (HD), generally three times a week. Many of these patients receive HD via a tunnel-type central venous catheter. For these patients, low catheter flow rates due to occlusion of the lumen thrombus still cause frequent complications and have been estimated to affect 3%-10% of all HD treatment periods and 87% of all catheters at some point prior to removal. (Moss et al., dm 尤/办叮Db; 12:492-8 (1988); Gibson and Mosquera, Ζ)ζ·α/ TValike/?/(10)ί 1991; 6:269-74 (1991); Suhocki^ A ^ Am J Kidney Dis, 28: 379-86 (1996)) The Kidney Dialysis Outcome Quality Initiative (KDOQI) Clinical Practice Guidelines for Vascular Access Abnormal catheter function was defined as "cannot obtain and maintain a blood flow rate outside the body (HD 300 mL/min) without significant prolonged hd treatment (National Kidney Foundation, K of vascular access) /DOQI Clinical Implementation Guide··2000 update. Am J Kidney Dis 37: SI37-81 (2001)). The KDOQI guidelines for the treatment of failed catheters recommend the use of thrombolytic agents as first-line therapy. Tupperase treatment of HD catheter work Abnormalities have been reported in (for example) Daeihagh et al. dm J Dis 36:75-9 (2000); Habowski et al. J Soc Nephrol 126382.doc -12- 200826981 11··185Α (2000) ; 'Mara et al. J dm #叩/2/^/·, 11:292A (2000); Roberts et al. t/Xm 11:195A (2000);

Zacharias et al. 27-33 (2000);

Hammes et al. J Soc 12:290A (2001) ; Spry and

Miller, Dial Transplant 30:10-2 (2001); Cairoli O.

Practical application: using t-PA (CathfloTM Activase®) overnight in catheter clearance on tunnel catheters used for hemodialysis. ^ (Proceedings of the 22nd Annual Conference on Dialysis) T2Lmpa. (FL) (March 4-6, 2002); Eyrich et al. Dm «/ do w P / mrm 59: 1437-40 (2002); Little and WalsheDz, 39:86-91 (2002); anti-OovAing special ruler. Nephrol Nurs J; 31:199-200 (2004). A dose of 1 to 2 mg of alteplase is administered in different volumes, with a residence time ranging from 20 minutes to 96 hours. Most of these studies have a small number of patients, use different dosing regimens, have a small amount of safety information, and have different efficacy definitions. Therefore, thrombolytic agents have not been studied in randomized, well-controlled clinical trials or their HD catheters approved for treatment of obstruction by the U.S. Food and Drug Administration (FDA). U.S. Patent Application Serial No. 10/697,142, filed on Oct. 30, 2003, the disclosure of which is incorporated herein by reference to the entire entire entire entire entire entire entire entire entire entire entire entire entire entire entire- Learn to collect. It is desirable to use a fibrin-specific plasminogen activator of an HD catheter that effectively and uniformly removes a pathological collection of fluids. For example, 126382.doc -13 - 200826981 allowed 1 than in 2003. The clinical application of higher doses of tenecteplase in US Patent Application #697142, filed on March 3, 1989, is required for HD catheter-guided blood collection. In particular, it is necessary to locally deliver blood to the lumen of the HD catheter during exposure, = lysogen W to provide remediation with suboptimal flow rate (4) = = risk of slimming-related adverse events associated with the use of the agent Minimal due to the time limit of the HD treatment period, there is a need for a stripping agent such as tenectease, "potency, high ▲ fibrin specificity and rapid dissolution of coagulation. In addition, fibrin is present in the prevention and removal of these rib devices. There is a continuing need for the fact that certain bacteria, in particular, have binding sites for retinoic proteins. ^ SUMMARY OF THE INVENTION Accordingly, the present invention is claimed as follows. In one embodiment of the invention herein, a recovery is provided in a mammal. A method of functioning a function of an abnormal hemodialysis catheter having a blood flow rate (BFR) of less than 3 〇〇 ml/min. m comprising locally administering a total dose of about 3 to 4 tenectease to all intraductal The chamber is allowed to allow for tenepase to remain in the catheter for about 1 hour to about 72 hours so that the flow rate of the catheter is no longer blocked. In one embodiment, during the 3 minute period prior to hemodialysis, at 25 The arterial dysfunction of mmHg has a BFR that is at least 25 mg/ml lower than the prescribed BFR. Tenapase is preferably in a solution of sterile water for injection or bacteriostatic water for injection. Preferably, it is retained in the catheter until the bfr of the catheter is increased beyond the BFR of the technology and tenecteplase and the increase is maintained for at least 48 hours. In its preferred embodiment, 126382.doc -14-200826981, tenecteplase The tenectease is administered to the intravitreous nevus in the sterile water for injection and/or at a total dose of about 4 mg, and preferably about 2 mg / 2 mL of the tenae f enzyme is administered to In two preferred embodiments, the tenecteplase is injected into the catheter for about an hour or as a length of more than about 1 hour to about 72 hours [preferred retention of about 2 to In about another 48 hours. In another preferred embodiment, the catheter is contacted with the solution for at least about 5 days to remove the fibrin-bound blood clot. In another preferred embodiment, the treatment is reproducible, ie Administration of tenecteplase over-time. The procedure is to be administered during the hemodialysis treatment period experienced by the mammal. Nipproxase. In another preferred embodiment, no retreatment is performed '(iv) Tenepase is administered as an initial dose only once or twice and subsequently administered as an extended indwelling dose. Optimally, tenecteplase It is administered only once, that is, as a dose. In other preferred aspects, the mammal undergoes hemodialysis after administration of tenecteplase. In other embodiments, the mammal is a human. Provided in a kit comprising a container comprising a solution comprising tenecteplase and a method for using the solution to restore the function of a functional abnormal hemodialysis catheter remaining in the mammal, wherein the guide has At less than 300 ml/min, the instructions direct the user to topically administer a total dose of about 3 to 4 mg of tenectease to all of the catheter lumens and allow tenectease to remain in the catheter for about one hour. It takes about 72 hours so that the flow rate of the catheter is no longer blocked. In another aspect, the invention relates to a method comprising the manufacture of a tenecteplase 'It catheter capable of restoring a functionally abnormal hemodialysis catheter left in a mammal with 126382.doc -15 - 200826981 It has a gold liquid (BFR) of less than one milliliter per minute. In another aspect, the invention relates to tenecteplase for use in restoring the function of a functionally abnormal hemodialysis catheter left in a mammal having a blood flow rate (bfr) of less than 3003⁄4 liters per minute. The invention herein provides for the use of tenectilase to treat hemodialysis that is blocked and becomes functionally abnormal (IV), in particular because of the pathological collection of *fibrin-containing fluids retained in the catheter. Abnormal function. [Embodiment] Definitions As used herein, "hemodialysis catheter, or, hd catheter" means generally, but not necessarily constructed of, a plastic polymer such as polyurethane, polyox, or other polymer. A dialysis catheter that is suitable for catheter-guided therapy (ie, delivering 4-study therapy) to deliver hemodialysis. The catheters herein are indwelling catheters, such as intravenous or arterial hemodialysis catheters, including their tunnel-type catheters. Preferably, the catheter is not an implantable type 4, a non-sleeve type catheter or a guide type guide. The HD catheter is preferably not implanted in the jugular vein. The HD catheter lumen preferably does not require reverse flow. The best is a sleeve type tunnel type HD catheter. As used herein, "Functional Abnormality" HD catheters are as recommended by the National Kidney Foundation for vascular access/DOQI clinical implementation guidelines: 2 year update. Am j

Kidney Dis 37: S137_81 (2〇〇1), a catheter that fails to achieve and maintains an extracorporeal blood flow rate of 126382.doc -16 - 200826981 (2300 mL/min) without significant prolonged hd treatment . In general, this functionally abnormal catheter typically has a BFR of less than 300 ml/min at a maximum negative arterial pressure of 250 mmHg. Preferably, the functionally-obstructed catheter exhibits a proven BFR equal to or greater than 300 mL/min during at least one HD treatment period 7 days prior to treatment by the methods herein. If the HD catheter has a sustainable BFR equal to or greater than 300 mL/min after the subject is reset, it is not functionally abnormal. Functionally-used catheters herein are preferably free of mechanical, non-thrombotic obstruction (e.g., kinking in a catheter or suture of a compressed catheter) or evidence of obstruction caused by a known fibrin sheath. The 'recovery function' means that HD is successfully performed at least once after treatment with tenecteplase, i.e., generally as the physician specifies, unobstructed and allows HD to proceed at a minimum flow rate. In general, this means that the function is restored when the catheter with the suboptimal flow rate is remedied and the BFR is restored to at least 300 ml/min. Although the first HD visit to the subject after treatment has been shown to show clinical success, the preferred subject demonstrates continuous catheter opening for a period of time after the first visit. The indication of functional recovery after HD success is the percentage increase from baseline at the end of the first visit to HD. Baseline BFR is the BFR measurement obtained to determine if the patient is suitable for treatment. In a preferred embodiment, tenecteplase is stopped in the patient's catheter for about one hour, the tenecteplase instillation is stopped and all patients experience complete HD. Next, BFR is typically measured during the last 30 minutes of HD to assess catheter function. In this preferred mode, it is considered that the BFR of 2300 mL/min at the end of HD is maintained at least for the last approximately 30 minutes and the subject with a BFR increase of 225 mL/min from baseline is successfully treated, and 126382.doc -17-200826981 Subjects with BFR 2300 mL/min and an increase of $25 mL/min from baseline BFR and subjects with BFR < 300 mL/min failed treatment. Similarly, in a preferred embodiment, such subjects have a urea reduction rate (URR) of at least about 65% as assessed by BUN measurements before and after HD at the first HD visit after treatment with tenecteplase. In another embodiment of functional recovery, after ten days of inoculation of the tenepase in the patient catheter, the patient underwent complete HD and BFR was measured during the last 30 minutes of HD to assess catheter function. At the end of the first visit HD, the subject with a BFR of "200 mL/min but < 300 mL/min obtained a prolonged indwelling time of the second dose instillation until the second visit (up to 72 hours) . At the beginning of the second visit, the extended indwelling dose of tenecteplase was withdrawn from the catheter and BFR was measured at the beginning of HD. The patient experienced full HD and again measured BFR during the last 30 minutes of HD. In this second preferred mode, it is considered that the BFR of 2300 mL/min continued to maintain at least the last approximately 30 minutes after the end of the first or second visit HD and the subject with a 225 mL/min increase from the baseline BFR was successfully treated. Patients who underwent HD catheter reocclusion (BFR < 300 mL/min) within 21 days of the first visit exited the initial course of treatment and entered the retreatment process, during which they received another dose of tenecteplase. After 1 hour of indwelling time, the patient experienced full HD and measured BFR during the last 30 minutes of HD. In this case, patients treated with BFR 2300 mL/min and increasing from baseline BFR <25 mL/min and subjects with BFR < 300 mL/min failed treatment and retreatment. • 'Subject' means that the drug is infused or dripped into the catheter. This generally means that the lumen of the catheter is flushed with tenecteplase. Allowing tenepase to "retain" means 126382.doc -18- 200826981 It is generally said that tenapopyrase remains in the catheter to perform its recovery flow rate. This π indwelling π means indwelling in the lumen. As used herein, 'pathology of a fibrin-rich fluid" refers to a collection of fibrin-containing aggregates that cause problems with hemodialysis catheters. The fluids may be from any source, including blood, cerebrospinal fluids, and fluids from the membrane, pleura, or pericardial cavity, and may be treated by thrombolytic drugs due to their high fibrin content. Because of this, this collection includes both the inner tube of the fluid and the non-vascular collection. This collection of fluids is contained in the hemodialysis catheter. This fluid is preferably pathological because it causes abnormal function of the hemodialysis catheter, thereby limiting the effectiveness of the ▲ liquid dialysis. Effective hemodialysis is hemodialysis that is functional for the subject being treated. For therapeutic purposes, the term "mammal," refers to any of the items classified as feeding (four): including but not limited to human 1 animal, zoo animal, dragon and domestic animal or farm animal, such as a dog, Cats, cows, sheep, pigs, horses and primates, such as monkeys or humans. Mammals are preferably humans. The mammals herein must require HD, and preferably must be prescribed (4) equal to or greater than 3 〇 0 ml / Minutes. At least two days before treatment, the mammal has preferably inserted its HD catheter. Eligible mammals also preferably use the same catheter on the same type and model of HD device '(d) at least 4 consecutive fistulas. Treatment period. Preferably, the treating mammal is capable of instilling a fluid in a volume necessary for instillation of tenectease into the HD catheter. The mammal may be an adult or juvenile mammal (e.g., a human being less than 18 years old), but is preferably an adult mammal. That is, a human at least 18 years old. As used herein, "therapeutic composition" or "composition," is defined as comprising 126382.doc -19-200826981 Nipzyme, sterile water for injection or for injection Sterile water and a carrier acceptable any optional pharmaceutically, such as minerals, proteins and other known to those skilled in the art of excipients. The tenecteplase is preferably in the form of a lyophilized powder reconstituted in one of these types of water. As used herein, "solution" refers to a soluble mixture of ingredients, including complete solvation of the ingredients.

As used herein, the term π tenecteplase, also known as TNK-tPA, or the TNKASEtm brand tissue plasminogen activator variant, is referred to as T103N, N117Q, K296A, H297A from Genentech, Inc. , t298 variant of R298A, R299A, wherein Thrl03 of wild type tPA is changed to Asn (T103N), Asnll7 of wild type tPA is changed to Gln (N117Q), and Lys-His-Arg-Arg of wild type tPA (SEQ) ID ΝΟ: 1) 296-299 was changed to Ala-Ala-Ala-Ala (SEQ ID NO: 2) (KHRR296_299AAAA). See the Background of the Invention section and U.S. Patent No. 5,612,029. As used herein, '1 sterile injection "Water" or "'SWFI" means the same substance as defined by the United States Pharmacopeia (USP), which is a bacteriostatic, antimicrobial or added buffer and is only used in single dose containers. A sterile, pyrogen-free preparation for the injection of water for diluting or dissolving the drug for injection. ''physiological saline'' means an aqueous solution containing 0.9% sodium sulphate. It is also known as a 0.9% sodium sulphide injection. Liquid USP, non-heparinized physiological saline. Generally The saline is clinically used as a diluent for the drug administered by injection and as a plasma substitute. 126382.doc •20-200826981 "Antibacterial water for injection" or "BWFI" means, for example, the United States Pharmacopoeia ( Usp) A mixture of water and other amounts of benzyl alcohol as defined in the present invention. Mode for carrying out the invention In the context of the invention, it is provided that the dysfunctional hemodialysis is retained in the mammal. A functional method of the catheter having a BFR of less than 300 ml/min. The method comprises topically administering a total dose of about 3 to 4 mg of tenectease to all of the catheter lumens (there are typically two lumens present in the catheter) The chamber) and the tenectease is allowed to remain in the catheter for about an hour to about 72 hours, so that the flow rate of the catheter is no longer blocked. Tenepase is preferably retained in the catheter until the catheter has increased the melting rate over the castane Pre-enzyme bribery and the increase is maintained for at least (9) hours. In other sputum I-like sputum in the nepase injection into the catheter for about ^ hours or as an extension of more than about 1 hour to about 72 hours. Good retention about 2 to about 48 hours. In another preferred embodiment, the catheter is contacted with the solution for at least about 5 days to remove the fibrin-bound blood clot. In other embodiments, tenecteplase can be administered - times or _ More than once and preferably only after the treatment period of hd. The mammal is preferably a human. The body is §, and the first time the patient is screened and the first dose of tenectease is used in the patient. After the enzyme treatment, hd was performed. Generally, it is based on the inclusion and exclusion criteria to screen qualified subjects. At the beginning of HD (usually within the first 30 minutes) BFR < 3 (10) mL / min (all subjects are preferably based on the maximum negative arterial of 250 mmHg to accept tenecteplase. Obtained to determine the study The leg of the appropriateness is measured as a baseline ship. In the embodiment, the human patient preferably receives a dose of about 1 mg/mL for 126382.doc -21 · 200826981 Nipzyme, wherein the dose is about 2 mg per lumen/ 2 mL, the total dose is about 4 mg. In other words, the patient preferably receives a dose of about 2 mg / 2 mL of tenecteplase, usually infused into each of the two lumens of the HD catheter. After 1 hour of indwelling time, tenecteplase was stopped and all subjects experienced full HD. Next, BFR was measured in the last 30 minutes of HD to assess catheter function. In this preferred mode, Subjects at 2300 mL/min BFR continued to maintain at least the last 30 minutes at the end of HD and subjects who received a 225 mL/min increase in baseline BFR were considered successful and considered BFR 2300 mL/min and increased from baseline BFR <25 mL/min Subjects and subjects with BFR < 300 mL/min failed treatment. In this case, the first time Subjects with a BFR of 2300 mL/min at the end of the HD are preferably no longer treated. If the BFR is still <300 mL/min at the beginning of the next HD treatment period, then 2 mL is preferred at that time. Mg) Tenepase treatment of BFR < 300 mL/min at the end of the first visit to HD. After 1 hour of indwelling time, it is preferred to stop tenecteplase and the subject experienced complete HD. BFR was again measured during the last 30 minutes to assess catheter function. In this preferred embodiment, two visits were made after exposure to the final tenectease (ie, one dose of tenecteplase was administered) BFR of the first and third visits of the second and third visits of the subjects receiving two doses of tenecteplase (in the first 30 minutes) The HD catheter opening of a successful subject treated with one or a second HD visit. In a second preferred embodiment, a human patient receives up to three doses of 126382.doc -22-200826981 Nipzyme. Subjects will receive one or two doses during the initial treatment, and eligible subjects who have undergone catheter reocclusion within 21 days of the first visit will receive Additional doses are part of the retreatment process. In particular, patients are screened and treated at the first visit to the initial treatment procedure. Qualified patients are screened based on inclusion and exclusion criteria. Treatment with tenecteplase at the beginning of HD (in Qualified subjects with a BFR of <300 mL/min (all BFR measurements based on a maximum negative arterial pressure of 250 mmHg) within the first 30 minutes. Baseline BFR is as defined above. The patient is administered with 2 mL (2 mg) of tenecteplase infused into each of the two lumens of the HD catheter. After 1 hour of indwelling time, tenecteplase was withdrawn and all patients underwent complete HD. BFR was measured during the last 30 minutes of HD to assess catheter function. Patients treated at 2300 mL/min BFR continued to maintain at least the last 30 minutes at the end of HD and increased from baseline BFR > 25 mL/min were considered successful. Subjects with BFR23 00 mL/min and from baseline BFR booster <25 mL/min and subjects with BFR < 300 mL/min failed treatment. Subjects with BFR < 200 mL/min or BFR 2300 mL/min and increased from baseline BFR < 25 mL/min at the end of the first visit HD were no longer treated. For subjects with a BFR of 2200 mL/min but <300 mL/min at the end of the first visit to HD, 2 mL (2 mg) of tenecteplase was injected into each lumen of the catheter. As part of the initial treatment process. The dose is left for an extended period of time until the second HD treatment period of the second visit (up to about 72 hours). Patients receiving a prolonged indwelling dose of tenectepase withdrawal their dose from the catheter at the beginning of the second visit, and then measured BFR at the beginning of HD (within the first 30 minutes). The subject experienced full HD and measured BFR again during the last 30 minutes of HD 126382.doc -23 - 200826981. Subjects who successfully treated at the 1st or 2nd visit and who had hd catheter reocclusion (BFR < 300 mL/min) within 21 days of the first visit stopped the initial treatment and entered the retreatment process, during the retreatment 2 mL (2 mg) of tenecteplase was again injected into each lumen, followed by! Hours of indwelling time (at the time of retreatment Visit 1). The BFR was measured at the beginning of each of the two visits after the final tenectease exposure (within the first 30 minutes) to assess the success of the treatment at the first or second visit or re-treatment of the first visit. The examiner's HD catheter is open. This second option was designed to assess the efficacy of continuously administering up to three tenecteplase doses to restore the function of a dysfunctional HD catheter, although the first option assessed the efficacy of consecutive doses of up to two doses. Thus, the invention herein encompasses not only administering a first dose of tenecteplase to a subject in need of restoration of the function of the hemodialysis catheter, but also administering a subsequent dose of tenecteplase in the same amount, but The amount can vary. For example, at the beginning of each HD treatment period, this is done one or several times. It can be treated or retreated as needed. HD can be viewed multiple times to ensure catheter opening and H success. Preferably, only the sputum is given ^ ^ Λ u / ^ ^ Β , one or two times (in the form of initial instillation or extended indwelling dose) and optimally only - times. Tenecteplase may be administered in the form of tenecteplase and a suitable form or bacteriostatic water for injection and a solution such as physiological saline solution. Although it can be: the stability of the composition is used to replace the enzyme, and the preparation method of the present invention is not suitable for the liquid mixture, but it is preferred to use the sterilized water for injection or the lyophilized powder of the tube. Prepared by rehydration in bacteriostatic water. Preferably, tenecteplase is administered to all of the catheter lumens, preferably both lumens, at a total dose of about 126382.doc •24-200826981 4 mg. Most preferably, tenecteplase is in sterile water for injection. The amount of tenecteplase provided is such that in a clinical or medical setting (such as, for example, the clinical and technical endpoints as set forth in the definitions above), the clot is dissolved in any obstructed catheter and the dysfunctional hemodialysis is otherwise restored. The amount of function of the catheter, but not exceeding the amount that would be a dangerous amount in the body to cause complications. Examples of major adverse events include intracranial hemorrhage (ICH), major bleeding, thrombotic events, thrombosis, catheter-related blood flu infection (CRBSI), and catheter-related complications, as well as any procedural requirements for adverse events in other procedures. Examples of secondary complication include, and do not require hematoma or any site and/or conservative treatment without any transfusion, excretion, or prolonged hospital stay for any particular treatment. According to the invention herein, the rehydration tenecteplase is formulated to a concentration of about 〇·, even l, mg/mL, preferably provided in one of the waters as described herein and optionally having one of the wounded waters. The lumen of the catheter is approximately U mg/2 mL to 2 g mL. Most preferably, tenecteplase is formulated to a concentration of about 1 call/shelf (to provide about 24 doses of 2 mg of tenecteplase per lumen).

Compositions suitable for clinical use in the use of Tenepase of the present invention include sterile aqueous solutions or sterile hydratable powders such as; More than 1? The formulation is obtained from water, preferably sterilized water for injection or tecnepase of dipeptone for injection. Also usually includes appropriate concentration

A combination of a buffer such as a arginine matrix to maintain a suitable pH' typically to 7. 5° additionally or additionally, a compound such as glycerol may be included in the formulation to help maintain shelf life. The formulation preferably comprises 126382.doc •25· 200826981 arginine, acid and emulsifier, such as polyethylene oxide sorbitan fatty acid ester, such as POLYSORBATE 20TM polyoxyethylene 20 sorbitan monolaurate , POLYSORBATE 80TM polyethylene sorbitan monooleate or POLYSORBATE 65TM polyoxyethylene 20 sorbitan tristearate, which in some embodiments coexists with lyophilized tenepase. In a preferred embodiment, a suitable amount of a pharmaceutically acceptable salt is also employed in the formulation to render the formulation isotonic. For example, a non-lyophilized sterile solution may optionally contain physiological saline. Tenecteplase is sterile and non-corrosive in a vial containing 52.5 mg of tenecteplase, 0.55 g of L-arginine, 0.17 g of phosphoric acid, 4.3 mg of POLYSORBATE 20TM polyoxyethylene 20 sorbitan monolaurate The lyophilized powder is commercially available in the form of a 10 mL syringe for sterile water USP for injection. Alternatively, tenecteplase can be supplied with a 10 mL syringe for bacteriostatic water for injection. The preferred water for injection herein is sterile water for injection. As an example of a suitable dosage form, a vial containing about 50 mg of tenecteplase and arginine, phosphoric acid and POLYSORBATEtm emulsifier is reconstituted with 50 mL of sterile water for injection. In another embodiment, to rehydrate the product, about 10 mL of the preservative-free sterile water for injection USP is mixed with the tenectepase powder under sterile conditions to produce a final concentration of about 1 mg/mL. Alternatively, tenecteplase is rehydrated to a concentration of about 1 mg/mL in BWFI (0.9%) with complete protein viability. The unused rehydred tenecteplase can be stored for up to 8 hours at room temperature (15-30 ° C) or 24 hours at freezing (2-8 ° C). 126382.doc -26- 200826981 In a vial configuration, the vial can contain 2 mg of nepeptidase in 2 mL of water for injection. Also included in this paper are small abandoned tiles with a weight and volume between about 丨mg and 4 mg and between about 1 mL and 4 mL. Since the procedure for the use of tenectease herein involves a total dose of about 2-3 mg for catheter clearance, the rehydrated tenecteplase can be easily frozen for subsequent use. Many organizations rehydrate and freeze smaller aliquots of tenecteplase (2 and 5 mL syringes) for future use to minimize waste and reduce costs.

It is recommended to visually check the precipitate of the solution after dilution and before administration. Tenepase does not contain preservatives and is theoretically susceptible to bacterial contamination and biochemical degradation when left at room temperature for more than 8 hours. Although the manufacturer recommends changing the solution after 24 hours, the drug should be physically and chemically stable for 24 hours. The tenecteplase can be dripped into the catheter for up to about an hour or about, or can be extended for longer than about i to about 72 hours, preferably from about 2 hours to 48 hours. . It may also take more than about η hours. Preferably, the amount is about i hours or up to about 24 hours. If the fibrin-bound blood clot is removed from the catheter, the catheter can be contacted with the solution for at least about 5 days, preferably about 6 to 15 days. The dinaphthine solution can be instilled or infused into the guide by any suitable technique. Based on the general knowledge of dosing agents in the literature, practitioners will be able to easily design methods for administering the solutions herein. ... can be hunted by a variety of assays known in the art, and the enzymes, as indicated in the definitions section above, such as the percentage increase from baseline BFR in the case of a dying of a swollen bundle, such as HD Before and after Β·measurement of the urea reduction rate, the success rate of HD treatment after indwelling agent retention. Physician 126382.doc -27- 200826981 The most appropriate catheter-based form should be used continually. The risk factors for adverse bleeding caused by tenetase are similar to those associated with alteplase, υκ and other plasminogen activators. Variables associated with unfavorable risks include increased tenecteplase dose, duration of infusion, adjuvant anti-blood stasis therapy (eg, heparin, aspirin (aspiriq or other antiplatelet agents as described herein), hypertension, Increased age, severity of ischemia, and female sex. Physicians should be aware of these risk factors and use appropriate t-alarms during treatment. If adverse bleeding occurs during infusion therapy, the tenectease should be terminated immediately and administered to the blood. The product (fresh frozen plasma or condensed protein) is reversed for low coagulability. It is preferred to administer the tenectease without any other active agent. However, the present invention includes injecting an active drug other than tenecteplase into the catheter. Examples of such adjuvants include blood diluents such as heparin and heparin analogs, including low molecular weight heparin such as tinzaparin (1), certoparin, parnaparin, Nadroparin, ardeparin, enoxaparin (LOVENOXTM5 Aventis Pharma, Bridgewater) , NJ), reviparin, liviparin and dalteparin

(dalteparin), warfarin (3_(a-acetonylbenzyl)_4_hydroxycoumarin or COUMADIN®) or aspirin; an anticoagulant such as tpA; a tPA variant such as reteplase; Urokinase; streptokinase; and venom plasmin (alfimeprase) 〇 undergo HD treatment and other drugs can be administered directly to the subject when administering tenecteplase. However, at this time, if no other thrombolytic agent is administered directly to the patient, 126382.doc -28 - 200826981 is preferred. : Hai and other adjuvants can be combined with tenepase or by: slaves and catheters by the usual route and amount. When tenecteplase is used together with - or a variety of other drugs, the dosage unit of the drug containing the other drugs in addition to tenepase is more than that of a, and when it is mixed with unfractloned heparin, The chymase is not phased and can be placed in the hall, and heparin should be administered by an independent method. Non-transparent thinner refers to a drug that does not matter and can be associated with reduced efficacy. The other molecules are provided or administered in appropriate amounts in an amount effective for the intended purpose, which is less than the amount used by the right non-tenectilase alone. Ben Mao also provides kits. In an embodiment, the kit comprises a container comprising a solution comprising at least tenecteplase (preferably for sterile water for injection or water for injection) and using the solution to recover and remain in the mammal , the instruction manual of the function of the abnormal blood hemodialysis catheter, which has a BFR of less than 300 ml/min. The instructions guide the user to administer a total dose of tenectacase of about 3 to 4 mg. To all catheter lumens and allowing tenepase to remain in the catheter for about 1 hour to about 72 hours, the flow rate is no longer blocked. The above is a description of the usage of the recovery method. The kit may also include instructions for re-investment. It may also contain another container having an adjuvant as an active ingredient as described above and a protocol for co-administration with an effective amount of a disk solution. Preferably, this embodiment is abciximab, eptifibatide (four) sin (four) cisplatin hydrochloride peak nhyd_hl. )), heparin or warfarin, a container for the instructions for co-administration with a dilute solution in an effective amount: 126382.doc -29- 200826981 These other instructions included in the kit generally include information about the agent 1 Information on the timing of dosing and other guidelines for the treatment of HD catheters. The container of tenecteplase can be in unit dose, bulk (e.g., multi-dose package) or sub-unit dose. In this kit, tenecteplase can be packaged in any convenient and suitable package. For example, if the tenecteplase is a freeze-dried formulation, it is preferred to use an ampoule or vial with a flexible stopper as a container so that it can be easily made by "sub/main shot/; Rehydration of the drug. Ampoules with inelastic, removable occlusion, Zeng (magic glass) or elastic plugs are most conveniently used for solutions of tenectease in Lu. In the latter case, the instruction manual also states that the contents of the vial are placed in a conduit for direct delivery. Various features and aspects of the present invention will be further described in the following examples: "The examples are provided to demonstrate how the skilled person operates in the context of the present invention" but it is not intended to serve as a vane to the invention in any way. Limitations. The disclosures of all references cited in this article are explicitly incorporated herein by reference. Example 1 The objectives of the study exemplified in the examples are compared with placebo controls. Efficacy and safety of the function of the HD catheter with abnormal enzyme recovery function. Abbreviation and definition list of terms

Adverse events Acute myocardial infarction Blood flow rate 126382.doc -30- 200826981 BUN Blood urea nitrogen CRBSI Catheter-related bloodstream infection CVA Central venous catheter CRF Case report form DMC Data Monitoring Committee EC Ethics Committee FDA US Food and Drug Administration GCP Good Clinical Standardize HD Hemodialysis ICH Intracranial Hemorrhage IND Research New Drug IRB Institutional Review Board IVRS Interactive Voice Response System KDOQI Kidney Dialysis Results Quality Guidance PCI Percutaneous Coronary Intervention QLab Quintiles Lab RT Retreatment SAE Serious Adverse Event SDV Source Distinction Verification SWFI Urine Urea Reduction Rate for Injecting Water Urine Urea Reduction Rate Study Objectives • Evaluate the efficacy of tenipepsin to improve dysfunctional hemodialysis (HD) catheter blood flow rate (BFR) after 1 hour of indwelling time compared to placebo • Evaluation of the safety study of tenepase in the treatment of subjects with dysfunctional HD catheters. This is a phase III, randomized, double-blind, 126382.doc -31 - 200826981 placebo control in approximately 40 centers in the United States. Research. Approximately 150 patients required HD and had a BFR < 300 mL/min and defined as an arterial pressure of -250 mmHg during the first 30 minutes of HD (or no more than 250 mmHg under the agency guidelines for maximum negative arterial pressure) Subjects with a functional abnormality HD catheter with a BFR of at least 25 mL/min will be enrolled in the study. Subjects will be classified into three categories from baseline BFR: 0-199 mL/min, 200-274 mL/min, and 275-299 mL/min. The number of enrollees participating in the 0-199 mL/min and 275-299 mL/min categories will be limited to the maximum 10% of the subjects in each category. Based on the regular HD time course of each subject and a follow-up visit, the study will consist of 3 to 4 visits corresponding to the continuous HD treatment period. Subjects will receive up to two study medications. After providing written informed consent (if available, and child informed consent), select eligible subjects based on inclusion and exclusion criteria at screening visits. At the discretion of the investigator, the screening visit and the first visit can be combined. At the first visit, at the beginning of HD (within the first 30 minutes), the arterial pressure at -250 mmHg (or at the maximum negative arterial pressure, BB, no more than 250 mmHg) was <300 mL/min and Eligible subjects with a BFR that is at least 25 mL/min lower than the prescribed BFR will be randomly assigned at a 1:1 ratio to receive tenecteplase or placebo. The BFR measurement obtained at the time of the first visit to determine the eligibility of the study was the baseline BFR. For the subject, 2 mL of study drug (i.e., 2 mg tenecteplase or placebo equivalent) was instilled into each of the two lumens of the HD catheter. After 1 hour of indwelling time, the study drug is withdrawn and all subjects will experience HD or experience HD as likely to the prescription. At the beginning of HD, every 30 minutes thereafter, BFR was measured 30 minutes before the end of HD and at the end of HD to assess catheter function and determine the treatment outcome of 126382.doc -32-200826981 for the first visit. Subjects with BFR < 300 mL/min at the start of the second visit with 2 mL (2 mg) open-labeled tenepase (no treatment results for the i-th visit). After 1 hour of indwelling time, the study drug is withdrawn and all subjects will experience HD or experience HD as possible to the extent possible. At the beginning of HD, every 30 minutes thereafter, bfr was measured 30 minutes before the end of HD and at the end of HD to evaluate catheter function and determine the treatment outcome of the second visit. By the second visit after the final study drug exposure (ie, the second and third visits of the subjects who received the study drug treatment, and received the second study)

Each of the 3rd and 4th visits of the drug-treated subject measured the BFR performing a follow-up evaluation of the HD catheter function at the beginning of the HD (within the first 30 minutes). For these assessments, the researcher M will increase the BFR in an attempt to reach a prescription BFR within the first % of minutes. If the HD catheter is removed at any time for any reason, no treatment will be provided and no efficacy assessment will be performed (ie bfr measurement or blood urea nitrogen [bun] y knife analysis). However, the subject will continue to undergo a safety assessment (ie, recording adverse events and accompanying drug and antibody tests). Subjects with symptomatic hypotension may not receive study medication. The second visit from the start of treatment to the completion of the final study drug exposure (ie, the third visit to the subject receiving one study drug treatment, and the subject who received 2 study drug treatments) In the case of the 4th visit, all adverse events of the subject will be recorded. All subjects will undergo an antibody test after 3 weeks of _36 days or after the study has terminated early. Efficacy Results Measurement 126382.doc -33 - 200826981 The treatment success of this study will be defined as follows: BFR2300 mL at arterial pressure in the range of 0 to -250 mmHg, at the end of HD and 30 (10) minutes before the end of HD /min and an increase of 225 mL/min from baseline BFR (wireless reversal). Subjects with a BFR of 2300 mL/min and an increase of <25 mL/min from baseline BFR, subjects with BFR < 300 mL/min, and subjects with a reversed pipeline will be considered treatment failure. If the investigator determines that the subject's hemodynamics has become unstable (blood pressure drop or heart rate change) and therefore requires a reduction in BFR, then BFR measurements must be recorded before BFR is reduced. Before hemodynamic instability occurs The BFR within 30 minutes will be used to determine the outcome of the treatment. For subjects who have successfully treated at the 1st or 2nd visit, the catheter function is defined as an artery in the range of 0 to -250 mmHg at follow-up visits. Depression at the beginning of the HD treatment period (within the first 30 minutes) BFR 2300 mL/min and an increase from baseline BFR 225 mL/min (wireless reversal). For these assessments, the researchers will increase BFR in an attempt to reach within the first 30 minutes. Prescription BFR. I Primary efficacy results are measured as follows: • Percentage of subjects on the first visit regarding BFR treatment success (as defined above); Secondary efficacy results are measured as follows: • For the first time Percentage of subjects who maintained catheter function at the 2nd and 3rd visits (as defined above) at the 2nd and 3rd visits. • Pre-treatment and post-HD BUN measurements were assessed at the first visit. 265% of urine Percentage of subjects with reduction rate (URR) • For those who did not receive open-label tenaplase at the second visit 126382.doc -34- 200826981, as assessed by BUN measurements before and after the second visit to HD Percentage of subjects with a URR of 265% • BFR changes from baseline at the end of the first visit to HD • Subject to the following types defined by changes in BFR from baseline at the end of the first visit to HD Percentage: <0 mL/min, 0-24 mL/min, 25-49 mL/min, 50·99 mL/min, 100-149 mL/min, and >150 mL/min for the second visit For subjects treated with open-labeled tenepase, the secondary efficacy outcome measures also include the following: • Percentage of subjects who were successful in BFR treatment (as defined above) at the 2nd visit • For Percentage of subjects who maintained catheter function at the 3rd and 4th visits (as defined above) at the 3rd and 4th visits • BFR changes from baseline at the end of the second visit to the HD • Percentage of subjects with a URR of 265% as assessed by BUN measurements before and after the second visit to HD • If you are visiting the third visit Percentage of subjects with the same URR of 65% evaluated by BUN measurements before and after HD belonged to the percentage of subjects of the following types defined by changes in BFR from baseline at the end of the second visit HD. · < The safety results of the primary safety results measured by mL/min, 0-24 mL/min, 25-49 mL/min, 50-99 mL/min, 100-149 mL/min, and 21 50 mL/min are as follows: 126382.doc -35- 200826981 From the initial study drug administration to the second visit to the starting rod target hemorrhage, major bleeding 'target adverse events (cranial reduction and guide 4 Wang Wang' positive, catheter-related blood flow α wood and guide The incidence of secondary safety results of the official-related complications is as follows: The second visit does not accept the incidence of #奈普酶*, from the enumerated) (4) adverse events (such as above)

For the subjects who received tenectease at the second visit, the incidence of target adverse events (such as the above two) from the start of the second visit to the fourth visit • from the initial study drug The incidence of gross adverse events and the incidence of all adverse events from the start of the second visit to the second visit: • From the second visit since the second visit to the subject who did not receive tenecteplase The incidence of serious adverse events and the incidence of all adverse events from the first visit to the third visit • For the subjects who received tenectease at the second visit, from the second visit to the fourth The incidence of serious adverse events and the incidence of all adverse events at the time of treatment • The incidence of positive anti-Tenapase antibody test in the subjects with negative baseline test Safety design Tenep I I approved for reduction Mortality associated with acute myocardial infarction (ΑΜί). Adverse events associated with systemic doses of 3〇-5〇 mg of natal ami are well described and consist primarily of bleeding complications including major bleeding events 126382.doc -36-200826981 and intracranial hemorrhage. In the function, when it was treated with a thrombolytic agent, the other side of the adverse event was catheter-related blood-sucking; k-plug event. Based on the clinical experience of the treatment of AMI and CATHFLO@ ACTIVASE (alteplase) in the treatment of y lip labiac abnormal CVA catheter, it is expected that any of the patients who are attributable to tenep or hemorrhage or blood test The event is likely to appear within 24 hours of treatment. The second visit from the start of the research and development of the drug to the final study of the drug was completed to record all adverse events. According to the study and treatment (the study design according to the rib catheter function recovery, the subject will receive up to two study drug treatments. The first treatment, at the ith visit will be tenip / placebo (two) all Subject, and the second treatment at the second visit will be given open-label tenectease to eligible subjects in each person-time when the drug is administered to the party for the tester will be 2 claws [research drug (ie 2 mg) Tenapopeptidase or placebo equivalent) was injected into each lumen of the HD catheter. Adjuvant therapy and clinical specification. From the first visit to the second visit after the final study drug exposure, the use of fibrin was prohibited. Dissolving agents (except for research drugs), warfarin (except for low-dose warfarin for prevention) and normal or low molecular weight heparin (except for heparin used only or for prevention). The second visit after the visit to the final study drug violent route, the subject taking piavix® (clopidogrel bisulfate) may not increase the dose. The subject may continue to accept the treatment at the discretion of the treating physician. Other drugs and targets for their pathology Treatment 126382.doc -37- 200826981 During the study (to the second visit after the final study drug exposure), the use of fibrinolytic agents (other than the study drug) is prohibited, but the subject can continue at the discretion of the treating physician. Subjects received medications and standard treatments for their condition. Statistical Methods Primary efficacy analysis calculates the percentage of subjects who achieved treatment success and provided a 95°/〇 confidence interval based on an accurate method. Comparison of treatment arms using the Cochran-Mantel-Haenszel test This percentage is (by baseline BFR classification: 0_ 199 mL/min, 200-274 mL/min, and 275-299 mL/min). Loss of data is for analysis purposes, and for any reason the study stops unrealized treatment. Subjects who are successful (as defined above) will be considered treatment failure. Determination of sample size The sample size of 1 50 subjects will provide a placebo response rate of 0.05 using the bilateral χ2 test at 0.05 significance level versus 5%. Detection of 25% of tenepase reaction rate > 90% of the assay power. Interim analysis of the period during which the Data Monitoring Committee (DMC) will conduct cumulative safety data Detailed study design This is a phase III, randomized, double-blind, placebo-controlled study that will be conducted at multiple centers. About 150, 16-year-old subjects who require HD and have a functionally abnormal HD catheter Subjects will be enrolled in the study. Subjects will be classified into three categories by baseline BFR: 0-199 mL/min, 200-274 mL/min 126382.doc -38-200826981 and 275-299 mL/min. The number of registered persons in the 0-199 mL/min and 275-299 mL/min categories will be limited to the maximum 10% of the subjects in each category. Based on the regular HD time course of each subject and a follow-up visit, the study consisted of 3 to 4 visits to each subject corresponding to the continuous HD treatment period. Subjects will receive up to two study medications. For the first treatment, tenectease or placebo was given to all subjects at the first visit, and the second treatment, at the second visit, the open-label tenecteplase was administered to eligible subjects. After providing written informed consent (if available, and child informed consent), select eligible subjects based on inclusion and exclusion criteria at screening visits. At the discretion of the investigator, the screening visit and the first visit can be combined. At the first visit, at the beginning of HD (within the first 30 minutes), with an arterial pressure of -250 mmHg (or no more than 25 0 mmHg under the mechanism of maximum negative arterial pressure), <300 mL/ Qualified subjects with min and BFR at least 25 mL/min lower than the prescribed BFR will be randomly assigned at a 1:1 ratio to receive tenecteplase or placebo. The BFR measurement obtained at the time of the first visit to determine the eligibility of the study was the baseline BFR. For the subject, 2 mL of study drug (i.e., 2 mg tenecteplase or placebo equivalent) was instilled into each of the two lumens of the HD catheter. After 1 hour of indwelling time, the study drug is withdrawn and all subjects will experience HD or experience HD as possible to the extent possible. At the beginning of HD, BFR was measured every 30 minutes, 30 minutes before the end of HD and at the end of HD to assess catheter function and determine the outcome of the first visit (as defined above). 2 mL (2 mg) open-labeled tenepase treatment at the start of the second visit 126382.doc -39- 200826981 BFR < 300 00 / min of the subject (no treatment results for the first visit) . After 1 hour of indwelling time, the study drug was withdrawn and the subject experienced HD or experienced HD to the extent possible, such as a prescription. At the beginning of HD, BFR was measured every 30 minutes thereafter, 30 minutes before the end of HD and at the end of HD to assess catheter function and determine the outcome of the second visit. By the second visit after the final study drug exposure (ie, the second and third visits to the subjects receiving one study drug treatment, and the third time of the subjects receiving two study drug treatments) And every 4th visit) The BFR performs a follow-up assessment of HD catheter function at the beginning of HD (the first 30 minutes). For these assessments, the investigator will increase the BFR in an attempt to reach a prescription BFR within the first 30 minutes. If the HD catheter is removed at any time for any reason, treatment will no longer be provided and no efficacy assessment will be performed (ie BFR measurement or blood urea nitrogen [BUN] analysis). However, the subject will continue to undergo a safety assessment (ie, recording adverse events and accompanying drug and antibody tests). Subjects with symptomatic hypotension may not receive study medication. The second visit from the start of treatment to the final study drug exposure (ie, the third visit to the subject receiving one study drug treatment, and for the subject receiving two study drug treatments) The fourth visit to the clinic will record adverse events for all subjects. All subjects will undergo an antibody test after 30-36 days of the first visit or after the study is terminated. This double-blind, placebo-controlled clinical trial was designed to evaluate the efficacy and safety of tenecteplase versus placebo in restoring dysfunctional HD catheters, and is based, at least in part, on CATHFLO® ACTIVASE® (Alte 126382. Doc -40- 200826981 The approved dose of enzyme and the experience of using alteplase in CVA and HD catheters. CATHFLO8ACTIVASE® (alteplase) is currently approved for the treatment of functionally abnormal CVA devices. In clinical trials, up to two 2 mg doses of alteplase (lower doses for subjects weighing < 30 kg) (120 minutes of indwelling time after each dose) are effective for restoring functional dysfunction CVA devices And safe.

ReΠno et al. TTzromZ? //aemwi 69(6): 841 (1993) Initial studies using human plasma demonstrated that teenopeptase has an equivalent efficacy with tPA (alteplase). In addition, Keyt et al. The corpse roc TVai/dead/SW. tASyi 91:3 670-4 (1994) demonstrates that tenectease has 82% of wild-type tPA activity with respect to plasma coagulation in the in vitro system. These results suggest that tenecteplase can have an alteplase activity of 82% to 00% for its ability to dissolve blood clotting; therefore, for each lumen, teniprase is present in the current study at a dose of 2 mg. (2 mL), withdrawn after 1 hour of indwelling time. It should be noted that the 2 mg dose is 15-25 times lower than the approved systemic dose of tenecteplase in the treatment of AMI. There was no systemic administration in this study. However, in the case where the lumen size of the catheter is unknown or less than the dose of tenecteplase (2 mL) specified in the study, there is a portion of the administered dose (ie, the difference between the volume of 2 mL and the lumen of the catheter) that enters the systemic circulation. possibility. The dosing regimen was not significantly different from the dosing regimen of a Cathflo Activase clinical trial (A2055g, A2065g, and A2404g) administered to a child subject weighing <30 kg by a dose equal to 110% of the lumen volume of the catheter. In the event that all 2 mg doses are not intended to be administered as an intravenous bolus injection, this would result in an expected maximum plasma concentration of 0.25 pg/mL. To be correct, the maximum predicted concentration of 2 mg alteplase is 0.58 126382.doc -41 · 200826981 pg/mL. In comparison, tenaplase, which is usually used in the 30 mg dose of AMI, will produce a maximum plasma concentration in the range of 5.9 to 7.5 pg/mL (average data from the TIMI 10A and 10B trials) (Cannon et al. 1997, 1998). Similarly, Tanswell et al. (1992) predicted that patients receiving 1 mg of alteplase via an accelerated infusion protocol achieved a maximum concentration of approximately 4 gg/mL. In comparison, endogenously produced tissue plasminogen activator levels have been reported to be in the range of 0.002 to 0.021 pg/mL. The basic principle of registration of BFR < 300 mL/min under arterial pressure of -250 mmHg is based on the recommendations in the KDOQI guidelines for vascular access to HD, suggesting that BFR at 2300 mL/min is unrestricted. Provide adequate dialysis in the case of prolonged HD treatment. Since BFR is directly related to negative arterial pressure, this study set arterial pressure in the range of 0 to -250 mmHg to maintain consistent conditions for BFR determination. In addition, the KDOQI guidelines recommend measuring BFR at an arterial pressure of -25 0 mmHg to determine catheter dysfunction. Efficacy Outcome Measurements Primary efficacy results were measured as follows: • Percentage of subjects on the first visit regarding BFR treatment success (as defined above). The secondary efficacy results were measured as follows: • Percentage of subjects who maintained catheter function at the 2nd and 3rd visits (as defined above) for the subjects who were successfully treated on the first visit • as in the first Percentage of subjects with >65% urea reduction rate (URR) assessed before and after treatment at the time of the second visit • For those who did not receive open-label tenaplase at the second visit 126382 .doc -42- 200826981 The percentage of subjects with a URR of >65% evaluated by the BUN measurement before and after the second visit to the HD. • The change in BFR from the baseline at the end of the first visit to the HD. Percentage of subjects of the following types defined by changes in BFR from baseline at the end of the first visit to HD: <〇mL/min, 0-24 mL/min, 25_49 mL/min, 50-99 mL/min 100-149 mL/min and 2150 mL/min For subjects who were treated with open-label tenaplase at the 2nd visit, the secondary efficacy outcome measures also included the following: • At the 2nd visit Percentage of subjects with successful BFR treatment (as defined above) • For successful treatment at the 2nd visit For example, the percentage of subjects who maintained catheter function at the 3rd and 4th visits (as defined above) • BFR changes from baseline at the end of the second visit to the HD • If the amount of BUN before and after the second visit to HD Percentage of subjects assessed to have a URR of 265% • Percentage of subjects with a URR of 265% as assessed by BUN measurements before and after the third visit to HD • BFR from the end of HD at the second visit Baseline changes define the percentage of subjects for each of the following types: <〇mL/min, 0-24 mL/min, 25-49 mL/min, 50-99 mL/min, 100-149 mL/min And 2150 mL/min safety results measurement primary safety results measured as follows: 126382.doc -43- 200826981 Since the initial investigation of drug administration to the second visit to start the standard adverse events (ICH, major bleeding, collateral events The incidence of secondary safety results for the incidence of gold-detection, catheter-related bloodstream infections [CRBSI] and catheter-related complications are as follows: = No need to receive tenectease at the second visit In terms of the target adverse events from the start of the second visit to the third visit (as listed above) Incidence

For subjects who received tenectease at the second visit, the incidence of target adverse events (as listed above) from the start of the second visit to the fourth visit was from the initial study drug to At the start of the second visit, the incidence of serious adverse events and the incidence of all adverse events were from the second visit to the third visitor who did not receive tenectease at the second visit. The incidence of serious adverse events and the incidence of all adverse events at the second visit were significantly unfavorable for the subjects who received tenectease at the second visit from the 2nd visit to the 苐* visit. Incidence of events and incidence of all adverse events. Prevalence of positive anti-Temperizide antibody test in subjects with negative baseline test. Safety design Tenepase was approved for reducing AMI-related mortality. . Full description of /, the body of the body using a dose of 30. 50 mg of tenepase in the treatment of AMI-related adverse events and it mainly consists of a major bleeding event and ICH bleeding complicated by 126382.doc -44- 200826981 syndrome. The elimination of tenecteplase from plasma is divided into two phases with an average initial half-life of 20-24 minutes and an average final half-life of 90-130 minutes (Modi et al. Journal of Clinical Pharmacology, 40: 508-515 (2000)). Although the incidence of bleeding complications in subjects with AMI treated with tenecteplase was quantified, the incidence of bleeding complications associated with lower tenecteplase doses used in this study was Limited information. The incidence of ICH and major bleeding due to tenecteplase is expected to be relatively low in this study due to the proposed low dose, minimal systemic exposure to tenecteplase and the clinical use of CATHFLO8 ACTIVASE® (alteplase) to date. Experimental experience indicating that ICH was not reported and only 3 of the 1432 subjects experienced major bleeding. Another adverse event associated with the use of thrombolytic agents to treat dysfunctional catheters was catheter-related thrombotic embolic events. This event can cause pulmonary embolism, which can be life-threatening depending on the size of the pulmonary embolism. Based on the experience of urinary enzymes and CATHFLO® ACTIVASE® (alteplase) in CVA catheters, the incidence of clinically significant cast events associated with the use of tenectease clearance catheters is expected to be low. Based on the clinical experience of tenaplase in the treatment of dysfunction of AMI and Cathflo Activase CVA catheters, it is expected that any potential bleeding or thrombotic events attributable to tenecteplase will likely occur within 24 hours of treatment. All adverse events were recorded from the start of the study treatment to the second visit after the final study drug exposure. All serious adverse events will be reported to Genentech within 48 hours, regardless of cause or treatment. The DMC will review the cumulative data of the tenepase catheter removal protocol at predetermined intervals and will be responsible for recommending the continued safety of the study to the sponsor based on the results of this review process. Subjects with dysfunctional HD catheters are suitable for this study and will be screened using the criteria provided herein. In assessing the efficacy and safety of intraperitoneal infusion of tenectease in the function of restoring dysfunctional HD catheters, subjects randomized to placebo will serve as a comparison group. The study was conducted in a randomized, double-blind, and placebo-controlled clinical trial to minimize any bias. This study was conducted in accordance with the U.S. FDA, INTERNATIONAL CONFERENCE ON HARMONISATION E6 GUIDELINE FOR GOOD CLINICAL PRACTICE (GCP) and any national requirements. Materials and Methods Subject Selection Subjects with a functionally abnormal HD catheter (as defined above) based on BFR during the first 30 minutes of HD were eligible for this study. About 1 50 subjects were enrolled in multiple study sites. Use the inclusion and exclusion criteria listed below to screen subjects. Subjects included in the standard must meet all of the following criteria to be eligible for inclusion in the study: • Can provide written informed consent and follow the study evaluation for the entire duration of the study • Age 216 • The investigator believes clinical stability is 126,382. Doc -46- 200826981

• Use a sleeve tunnel type HD catheter with a BFR of <300 mL/min at a maximum negative arterial pressure of 250 mmHg, but with a proven 2300 mL/min during at least one HD treatment period 7 days prior to the first visit. BFR • HD BFR 2300 mL/min • Under arterial pressure of -25 0 mmHg (or no more than 250 mmHg under the mechanism of maximum negative arterial pressure) with baseline BFR of <300 mL/min (pre-HD During the 30 minute period (use the catheter line in the usual direction before any reversal of the line; see section 3.1.2.a) • Baseline BFR (during 30 minutes prior to HD) is at least 25 mL/min lower than the prescription BFR • For example, HD Subjects with a prescription BFR of 300 mL/min must have a BFR of $27.5 mL/min to enter the study. • At an arterial pressure ranging from 0 to -250 mmHg in at least one HD treatment period 7 days prior to the first visit, BFR proved to be 2300 mL/min (using a catheter line in the usual direction) • Expect at least 4 consecutive HD treatments Use the same catheter on the same type and model of HD device • Volume infusion fluid necessary to inject the study drug into the HD catheter (see dose, administration, and storage section below) Exclusion criteria meet any of the following criteria Subjects will be excluded from the study: • HD catheter with a BFR of 2300 mL/min after the subject was reset • HD catheter insertion before screening • 2 days • Mechanical, non-thrombotic origin of HD catheter dysfunction (eg catheter 126382.doc -47- 200826981 ket or compression catheter suture) or evidence of dysfunction by known fibrin slightly μ • implanted in the subclavian vein using implantable 埠• HD catheter • It is expected during the study to use a catheter for treatment, any other type of procedure (ie not HD) or to be treated in this study before the study or any of the tenectilase catheters. Use any research drug or green test within 28 days. Use fibrinolytic agent (such as azinase, tenecteplase, reteplase or urokinase) within 7 days before the first visit. Knowing pregnancy or breastfeeding • HD catheter with known or suspected infection • History of any intracranial hemorrhage, aneurysm or arteriovenous malformation • Use of any heparin (ordinary or low molecular weight) within 24 hours prior to the third visit The first person outside the heparin during HD or for prevention (eg heparin lock) uses warfarin within 0刖7 days, starting or increasing Plavix within 7 days before the first visit in addition to the low-dose warfarin for prevention. @(克罗比多氢盐) Dosage • In the case of erythropoietin, hemoglobin >13 5 Laboratory tests for hemoglobin content must be performed within 30 days prior to screening. 0 Platelet count <75,000/μί Laboratory tests for platelet count must be performed at 126382.doc -48-200826981 within 30 days prior to screening. • The investigator believes that a high risk of bleeding events or embolic complications (ie, recent pulmonary embolism, deep vein thrombosis, endarterectomy, or clinically significant right-to-left shunt) or a known condition with significant risk of bleeding • BFR < 300 mL/min due to symptomatic hypotension • Researchers believe uncontrolled hypertension (eg systolic blood pressure > 185 mmHg and diastolic blood pressure > 110 mmHg) • Known for tenecteplase or formulation A method for hypersensitivity treatment dispensing and blind testing for any component This is a double-blind, placebo-controlled study. Subjects will be classified into three categories by baseline BFR, 0-199 mL/min, 200-274 mL/min, and 275-299 mL/min. The number of registrations in the 0-199 mL/min and 275-299 mL/min categories will be limited to the maximum 10% of the subjects in each category. Within each category, stratified, dynamic algorithms were performed, performed via the Interactive Voice Response System (IVRS), and subjects were randomized to the tenectease or placebo group at a 1:1 ratio. Formulations Tenepopzyme and placebo were provided in disposable, 6 CC glass vials with DAIKYOTM stoppers and easy-to-open aluminum covers. Tenecteplase is provided as a sterile, lyophilized formulation containing 2 mg of protein having the following excipient specifications: 104.4 mg L-arginine, 32 mg phosphoric acid, and 0.8 mg polysorbate 20. A placebo is provided in the form of a sterile, lyophilized formulation having the same excipient as the tenecteplase without the active ingredient. Diluent is sterile water for injection USP/EP (SWFI) 〇126382.doc -49- 200826981 Dosage, administration and storage According to the recovery of HD catheter function as described in section 3.1, the subject receives up to two study drugs. treatment. At each treatment, 2 mL of study drug (i.e., 2 mg tenecteplase or placebo equivalent) was instilled into each lumen of the HD catheter for the subject. If at any time the hd \guide is removed for any reason, no further treatment is provided. Subjects with symptomatic hypotension * may not receive study medication. (Reconstitute each vial of lyophilized tenepase or placebo immediately before use with 2·2 mL of SWFI. Use sterile technique to direct SWFI directly into the lyophilized block of study drug and gently vortex the vial until the content The substance dissolves. It does not vibrate. The concentration of tenectease in the obtained solution is 1 mg/mL. It is not abnormal to slightly foam after rehydration; if the vial is allowed to stand quietly for a few minutes, any large bubbles will be dispersed. If the study drug is not used immediately, the solution must be stored at 2. 〇8 (: (36. 46 卞) and used within 8 hours of rehydration. Discard any unused solution. ϋ Only before administration of study drug , withdraw any fluid from the lumen of the hd catheter and attempt to flush with saline. For administration of the dose, 2 mL of reconstituted research drug should be drawn into a single 1 sputum using aseptic technique. According to the guidelines of the institution, the solution is subsequently It should be dripped into the lumen of a HD catheter. The remaining volume of the catheter should be filled with normal saline. Repeat for the second lumen. Store the study drug at 2°〇8 (36卞-46卞). Vial. Do not store any preparations will be Use of the unused portion of the vial not used vials provided more than tenentech or failure to extend the expiration date of the study drug on file. Partially used vial, the vial was empty and did not return to the rehydration 126382.doc -50- 200826981

Genentech 剂量 Dose change No dose change is allowed. Companion and exclusion therapy = face is not allowed to receive any intravenous therapy or the study drug is in a blood sample via a HD catheter. It can only be obtained by using intravenous therapy && From the first visit to the final music violent road section 2: owing to the completion of the visit (ie, for the subject receiving sputum treatment, the third visit, and for the subject receiving the two treatments 'The 4th visit, prohibiting the use of fibrin: Decomposing agents (other than research drugs), warfarin (except for low-dose Chinese or low-molecular-weight heparin for prevention (except during HD or for prevention) From the first visit of the younger brother to the second time after the final study drug exposure, Yuan Cheng, the subject taking the clinopyrine sulfate may not increase the pot. The examiner may continue to receive the vote at the discretion of the treating physician. With the drug and standard treatment. The disease study evaluation and = will be composed of the continuous TM treatment period corresponding to each subject in the follow-up treatment. At the discretion of the researcher, the screening of the ridge and the first visit can be combined Subjects will receive up to two studies of sputum: 庵 first treatment, in the flute] 钔人研九乐物治疗. There are subjects, 广? (4) will be tenipeptin or * consolation to give it The enzyme treatment will be open at the 2nd visit and will be in the 2nd and second Subjects who received the study drug treatment were followed by a follow-up assessment of hd catheter function. 126382.doc -51 - 200826981 Subjects treated with two study medications will be in the 3rd and 4th The visit experienced a follow-up assessment of HD catheter function. ^ 3 visits (up to 36) days later or after the study was terminated early, all subjects will return to follow up. If at any time the HD catheter is removed for any reason, it will not The treatment is provided and the efficacy assessment will no longer be performed (also (4) FR measurement or _ analysis, and: 'The subject will continue to undergo a safety assessment (ie, recording adverse events and accompanying drug and antibody tests).

A laboratory kit and instructions for collecting BUN and anti-tenopeptidase antibody samples are provided by the Central Laboratory Quintiles Laboratory (QLab). All samples will be processed in situ and shipped to QLab. QLab will perform a coffee analysis, calculate the samples and ship the antibody samples to Genentech for testing. Screening visits At the discretion of the investigator, any or (four) (four) assessments can be performed at the first visit (before randomization). Written informed consent/consent must be passed before any research-specific assessment or procedure is performed. The following screening assessments and procedures will be conducted: • Written informed consent/consent • Review of inclusion and exclusion criteria • Demographic data including the date of birth, gender and race/racial • physical examination and medical history' including two recent Ship value (historical baseline) Right on the screen, if the medical examination is not indicated at 0, the historical body can be used for the check, and the restriction is made within 7 days before the screening. • Signs of birth τ include blood pressure, respiration rate, temperature and pulse (detailed after 126382.doc -52- 200826981) • Weight (detailed before and after HD) • If before screening 30 Within the day of laboratory testing to confirm eligibility, blood samples are taken to determine hemoglobin content (if the subject is for erythropoietin) and platelet counts • Accompanying drugs • HD catheter history and information will be recorded for HD catheterization and Finally, the information on the date of the function (BFR23〇〇mL/min) of the HD catheter is known. The size, type, volume, brand (if known) and placement of the HD catheter lumen are also recorded. HD prescription The first visit The first visit must be made within 7 days after screening (as described above, the screening visit and the first visit can be combined at the discretion of the investigator). At the beginning of the first visit, the following procedures should be performed to confirm eligibility: • Review of inclusion and exclusion criteria • Review of accompanying medications and medical history (to ensure no change since screening), including prior to the first visit Use of fibrinolytic agent, warfarin and crotidol bisulfate in 7 days and use of heparin within 24 hours before the first visit (see section 4.1.3) • HD prescription • HD, start with prescription • Baseline BFR, measured at the beginning of HD (within the first 30 minutes) to confirm HD catheter dysfunction 126382.doc -53- 200826981 at -250 mmHg arterial pressure (or institutional criteria for maximum negative arterial pressure, no more than 250 mmHg) Subjects with <300 mL/min and BFR at least 25 mL/min lower than the prescribed BFR (using a catheter line in the usual direction) are suitable for this study; all other subjects are not suitable. The baseline BFR was recorded the first time the subject was determined to be suitable for the study (i.e., when the arterial pressure reached -250 mmHg). If dialysis with a reverse line has been attempted (i.e., prior to selection of study medication), BFR must be recorded prior to online reversal and used as a baseline value. Subjects who are unable to measure BFR due to total obstruction (ie, no blood draw function) should be considered to have a baseline BFR of 0 mL/min. A qualified subject will interrupt their HD. Unqualified subjects may complete their HD treatment period and should be registered as a screening failure. After eligibility was confirmed, subjects were randomized using IVRS and blinded to study the drug kit. The following assessments and procedures were also performed at the first visit • Collecting blood samples for serum anti-Tenapase antibody test before study drug treatment • Collecting blood samples for BUN analysis before treatment with the study drug • Restoring HD pre-blind study drug administration The study drug was administered as described in section 4.3.2 and allowed to stand quietly in the subject's HD catheter (two lumens) for 1 hour. After leaving for 1 hour, the study drug was withdrawn. Subjects with symptomatic hypotension may not receive study medication. • Recover and perform HD as scheduled and resume and execute HD to the possible extent 126382.doc -54- 200826981 • At the beginning of HD, every 30 minutes thereafter, measure BFR 30 minutes before the end of HD and end of HD to determine treatment Results (as defined in Section 3.1.2.b) If it is necessary to reverse the catheter line dialysis, the BFR measurements are recorded prior to reversal and other BFR measurements will not be recorded during this HD treatment period. If the investigator determines that the subject needs to reduce his BFR due to hemodynamic instability, the BFR measurement is recorded before the BFR is lowered. Subsequent BFRs will continue to be recorded as scheduled. • Collect blood samples for BUN analysis after completion of HD If the catheter line has been reversed, no blood samples will be taken. • Adverse events and changes in accompanying medications during this visit The study began to monitor adverse events after initiation. The second visit will perform the following assessments and procedures at the second visit: • Adverse events and accompanying medication changes after the last visit • HD prescription • Blood samples collected for BUN analysis before HD • HD, starting with prescription • BFR, measured at the beginning of HD (within the first 30 minutes) to assess HD catheter function For these assessments, the investigator will increase BFR in an attempt to reach a prescription BFR within the first 30 minutes. Subjects with a BFR of 2300 mL/min at the start of HD will complete their HD treatment period. Subjects with BFR < 3 00 mL/min will have their HD interrupted and perform the following assessments and procedures: • Open-labeled tenectease administration 126382.doc -55- 200826981 Submitting Tina as described in Section 4.3.2 The enzyme was allowed to stand quietly in the subject's HD catheter (two lumens) for one hour. After staying for 1 hour, the tenecteplase was extracted. Subjects with symptomatic hypotension may not receive study medication. - • Recover and perform HD as required and resume and perform HD to the extent possible' • At the beginning of 110, every 30 minutes thereafter, measure BFR 3 minutes before the end of HD and end at the end of ^ to determine the treatment outcome (eg 3 ·丨2 b is defined) ^........, if it is necessary to reverse the catheter line dialysis, the BFR measurement is recorded before the reversal and other bfr quantities will not be recorded during this HD treatment period. Measured value. If the investigator determines that the subject needs to reduce his BFR due to hemodynamic instability, the BFR measurement is recorded before the BFR is lowered. Subsequent bfr will continue to be recorded as scheduled. All subjects after HD completion (regardless of whether they receive open-label tenaplase) will perform the following assessments and procedures: • Blood samples for BUN analysis (if the catheter line has been reversed, no blood samples will be taken. • During this visit Adverse events and changes in accompanying medications The third visit to the third visit to perform the following assessments and procedures: Final - adverse events after the person's shirt and changes in accompanying medications • HD prescriptions • For the second visit only Subjects receiving open-labeled tenecteplase: HD-collected blood samples for bun analysis • Performed 110 as prescribed or performed HD to the possible extent 126382.doc •56- 200826981 • BFR, at the beginning of HD (before Within 30 minutes) to assess the function for 4, 4, the researchers will increase the BFR in an attempt to achieve a prescription BFR within the first minute. • For subjects who received only open-label tenaplase at the 2nd visit: Blood samples collected for bun analysis after HD completion will not be collected if the catheter line has been reversed. • Unfavorable events during this visit and changes in accompanying medications The fourth visit is only open for the second visit. The subjects who received the tenectease require a fourth visit. The following study assessments and procedures will be performed on their subjects: • Adverse events after the last visit and changes in accompanying medications • For the second time only Subjects who have successfully treated the treatment: BFR is measured at the beginning of HD (within the first 30 minutes) to evaluate HD catheter function • HD prescription • Perform HD or perform HD to the extent possible • BFR 'At the beginning of HD ( Measurements to assess hd catheter function month b ° For these assessments, the investigator will increase bfr in an attempt to achieve a prescription BFR change within the first 3 minutes. • Adverse events during this visit and accompanying medications Changes were performed on the 30th day/early termination. The blood of all subjects was taken for anti-Tenapase antibody test on the 30th day after the first visit (up to 36 days) or after the study was terminated early. 126382.doc -57- 200826981. If the study is finally administered, the patient will be recorded at the early termination and the information about the catheter status will be collected. The adverse event will occur before the second visit. Subject to inspection At any time, the person has the right to withdraw from the study. The researcher is entitled to withdraw from the talented person for any reason of the subject's maximum I 丨 if if , , , , , , , 。 。 。 。 。 。 。 。 。 。 。 。 。 。 * * * Identifying a disease, an adverse event, or a deteriorating condition. Genentech reserves the right to suspend the program, control the original, or restrict or terminate the decision of the Nine-Year for any reason or any reason. The right to withdraw from the subject. The study ceases enentech's right to terminate the study at any time. Reasons for terminating the study may include, but are not limited to, the following: • (4) or other studies where the incidence or severity of adverse conditions indicates The examiner has a potential health hazard. • The registration of the subject is unsatisfactory. • Data records are inaccurate or incomplete. Statistical methods j machine = and all subjects treated with research drugs will be included in the efficacy and An Wang knife analysis. The efficacy analysis will be based on the assigned treatment of the subject, and the analysis will be based on the actual treatment of the cross-checker. All hypothesis tests will be performed at a 0.05 significance level, but not for multiple endpoints. Full details of the statistical analysis will be included in the statistical analysis design. The research implementation is divided into two categories: 126382.doc -58- 200826981 The treatment arm will summarize the registration number, the violation, the study stop and stop the original subject deployment form. The number of tinepops administered, the main program. The treatment group will be treated at each study visit.

U Analytical treatment group comparability summarizes demographic and basic characteristics, such as age, gender, race, weight, catheter type, and baseline bfr, by using the mean and standard deviation or median and the range of continuous variables and the proportion of categorical variables. . Statistical tests for significant differences between treatment arms will not be performed. Efficacy analysis a. Primary efficacy outcome measurement The primary efficacy outcome measure is the percentage of subjects who were successful in BFR treatment (as defined previously) at the third visit. Those who believe that the study is stopped before the completion of HD or the primary outcome measure cannot be evaluated in other ways are related to the failure of the primary outcome test. Calculate the percentage of subjects who achieved successful treatment and base them on the 953⁄4 confidence interval. This percentage between treatment arms was compared using the c〇chran-Mantel-Haenszel test (classified by baseline BFr: 0-199 mL/min, 200-274 mL/min, and 275-299 mL/min). Sensitivity analysis was performed to assess the robustness of the primary outcome of the alternative missing data method, which included a complete case analysis and a final observation carried forward (LOCF) estimate. b. Secondary efficacy outcome measures For subjects who were successful in the first visit, the percentage of subjects who maintained catheter function at the 2nd and 3rd visits (as defined previously) was similar Analysis of primary efficacy outcome measurements. The percentage of subjects who maintained 126382.doc -59- 200826981 catheter function at each visit was calculated and a 95% confidence interval was provided based on an accurate method. Successful treatment at the first visit, discontinuation of the study before the completion of the 2nd and 3rd visits, or a BFR unpredictable at the time of such visit will be considered as a failure to measure the secondary outcome measurement. At the first visit, the URR is calculated as follows: (BUN before treatment) - (BUN after HD) (BUN before treatment) At all other visits, URR is calculated as follows: (BUN after BUNHHD before HD) (BUN before HD) For each of the 1, 2, and 3 visits, the percentage of subjects with a URR of 265% was calculated and a 95% exact confidence interval was provided. At the first visit, the Cochran-Mantel-Haenszel test was used to classify the percentage of subjects with a 265% URR between treatment groups by dialysis time point and dialysis duration. Subjects who discontinue the study prematurely or who are not valuable by URR will be considered to have a <65% URR for this result. The treatment group will summarize the mean change in BFR from baseline at the end of the first visit to HD and provide a 95% exact confidence interval. Treatment comparisons were performed by baseline BFR categorization using the Cochran-Mantel-Haenszel test. The following change categories were also used to analyze changes in BFR from baseline at the end of the first visit to HD.·· <0 mL/min, 0-24 mL/min, 25-49 mL/min, 50-99 mL/min, 100 -149 mL/min and 2150 mL/min. For these results, subjects who missed BFR data have values estimated using the LOCF method. Use the alternative estimation method (described in Statistical Analysis Design) to evaluate the robustness of the results 126382.doc -60- 200826981. An analysis similar to the primary and secondary outcome measurements described above was performed on the measurement of the results of the open labeling of the tenectilase (see section 3 3) at the second visit. For these analyses, no comparison of treatment groups was obtained, as all treated subjects received open-label tenaplase. e•Subgroup analysis

Provide estimates of the primary and secondary efficacy outcome measures and confidence intervals and key safety outcomes for the following subgroups: • Age: <18, 18-65, > 65 years • Gender: Male, Female • Baseline BFR : (M99 mL/min, 200-247 mL/min, 275_299 mL/min Safety Analysis Using the MedDRA Dictionary, the literal description of the treatment emergency μ event was converted into the car owner term and body system terminology. Event_Overview of the unfavorable events from the initial study drug administration until the second visit. A similar overview of the body system, high-level injections and preferred terms for all adverse events and serious adverse events at this same interval. Summary of the comparison between the safety of the testosterone and the placebo-treated subjects. For the subjects who continued to receive tenectease in the second visit, the ancient == category summary from the second time From the start of the visit to the 4th time: the completion of the W adverse event. The body system, high-level terminology and better for all adverse events and serious events of the patients at the same interval 126382.doc 2008269 81 A similar overview of the terminology. For the subjects who were not treated with tenectease at the second visit, the classification of adverse adverse events from the beginning of the second visit to the third was completed. Adverse events. A similar overview of all of the patients' (4) events and serious (4) events (4), high-level terms, and preferred \ terms are generated at the same interval. - Baseline antibody status and tenecteplase exposure status The results of the tenecteplase antibody test during the tracing period were tabulated. The data for the anti-leakage was analyzed (4), the study was stopped for any reason, the treatment was not achieved (as defined above) and the BFR increased from the baseline &gt The subject of ;25 will be treated as a treatment failure. The sample size is determined. The primary efficacy result is measured as the success of the treatment (as defined above). The number of surgeons is 155. The subject is enrolled and 1 · _ The 1 ratio was randomized to the I: Nipase group or the placebo group. The sample size was provided using a bilateral χ2 test at a steep level relative to 5% of the placebo success rate detection of 25%, Nip Enzyme treatment success rate > 90% verification The mid-term analysis forms the DMC and is responsible for the periodic review of cumulative safety data during the study period. The DMC operates independently of the sponsor and Quintiles and consists of clinicians and biostatisticians with appropriate (7) expertise. Interval review of cumulative safety data for the tenectease catheter removal program, the study of the Spear G Gluconium abnormal CVA& HD catheter (study N3698g, Μ% LV, 126382.doc -62-200826981 N3700g and N3701g) Based on the results of the review process, the project will be responsible for making recommendations to the sponsor regarding the continued safety of the study. The dmC2 specific guidelines and operational procedures will be summarized in the DMC regulations. Safety Assessment The safety assessment will consist of monitoring and recording adverse events (AE) and severe adverse events (SAE) including target AE. Adverse Events Regardless of attribution, AE is any undesired, undesired sign, symptom or disease associated with an intervention applied using a research (medical) product or other protocol. It includes the following:

• AEs that were not observed in the subject during the AE reporting period specified in the protocol • Complications that occur as a result of the intervention required by the protocol (eg, invasive procedures, such as biopsy) • At the program Pre-existing medical conditions (except for the condition being studied) that are determined by the investigator to deteriorate or change in nature during the prescribed AE reporting period. Severe adverse events if the AE meets the following criteria, It is classified as SAE: it causes death (ie, AE actually causes or causes death). It is life-threatening (ie, the investigator believes that AE puts the subject at risk of direct death. It does not include AEs that can lead to death if they appear in more severe forms). 126382.doc •63 - 200826981 • It requires or extends hospitalization for inpatients. • It results in persistent or apparent disability/inability (ie, AL. The ability to cause the subject to perform normal life functions is generally disrupted). • It causes congenital anomalies/congenital defects in newborns/babies born to mothers exposed to the study drug. • It is a significant medical event considered by the investigator based on medical judgment (for example, the subject may be compromised or may require medical/surgical intervention to prevent one of the above listed results from occurring). All AEs that do not meet any of these serious criteria should be considered non-critical AEs. The terms "severe" and ", serious", (,, (8) cap,,) are not synonymous. Severity (or intensity) refers to the level of a particular AE, such as mild (grade 〇, medium Degree (level 2) or severe (level 3) myocardial infarction.,, severe, for management ambiguity (see previously defined) and based on the usual examination of events related to the life or function of the subject& Severity (non-severity) serves as a guide for defining management reporting responsibilities from sponsor to appropriate authority. Fields should independently assess severity and severity for CRF records AE and SAE. Adverse event

In particular, events that cause specific attention (target AE) and include the following events: • Computerized tomography imaging or magnetic resonance imaging of ICH major bleeding, defined as severe blood loss (>5 mL/kg), requiring blood transfusion or Caused by low jk pressure loss of blood 126382.doc -64 - 200826981 2 plug 'defined as any 'severe, embolic events, including lung events, pulse events (9) such as stroke, peripheral embolism or major organ thrombosis cholesterol plaque: 'Conduit-related venous thrombosis, which is defined as pain, swelling, and/or localization of limbs caused by radiography, imaging (eg, ultrasound, a radiograph, or magnetic resonance) in the upper or lower extremity arteries or veins. The ischemic thrombus CRBSI is further classified as follows: Clear type: no other significant source of infection. Semi-quantitative cultures from the tip of the catheter in the subject (each catheter fragment > 15 colony forming units) and from the periphery or catheter The same organism of the blood sample may have a monthly b-type. In the symptomatic subject without other obvious source of infection, the blood culture confirms the infection but the catheter tip does not confirm the infection (or catheter) The situation confirmed by the tip, but the blood culture was not confirmed), (4) or the case of not removing the catheter. 'The symptoms of antipyretic symptoms after antibiotic treatment. Potential symptoms: There is no other bloodstream infection in the symptomatic subjects without other obvious sources of infection. Symptoms of catheter-related complications after catheterization or removal of catheters • Catheter-related complications defined as catheter rupture, indwelling venous perforation, or surgical intervention during irrigation or infusion of medication (eg Bleeding at the catheter insertion site of suture or gauze. If the target meets the criteria for severe adverse effects as described above, it should be classified as SAE and should be reported. ' ~ Adverse event reporting period 126382.doc -65- 200826981 All study periods for which AE and SAE must be recorded begin at the start of the study treatment and the second visit after completion of the final study treatment (ie, for the subject receiving a study drug treatment, the third time) Visiting the doctor; and for the subject who received two study medications, the fourth visit) or when the subject stopped the study (any prior). Assessment of events In addition to the follow-up visit on the 30th day, the investigators assessed the presence of AEs and SAEs at the time of assessment of each subject during the study period. All were discovered by the investigator during the inquiry or by the investigator. AEs and SAEs detected via physical examinations, laboratory tests or other methods will be recorded in the medical records of the subject and on the appropriate AE or SAE CRF page. Each record AE or S AE is in its duration (ie Start and end periods), severity (see Table 1), (if applicable) management severity criteria, suspected relationship with study medications (see rules below), and actions taken to describe them. Table 1 Levels of adverse events (severity) Scale severity describes mild temporary or mild discomfort (<48 hours); does not interfere with the subject's normal activities; does not require medical intervention/therapy moderate to moderate to moderate disturbance of the subject's regular activities; The need for minimal medical intervention/therapeutic severity causes considerable disruption to the subject's abnormity; medical intervention/therapy is required; possible hospitalization notes: no severity Some events can also meet management severity criteria. See the SAE definition in this article. 126382.doc -66- 200826981 To ensure the AE and sae cause assessment, the investigator should use the following general guidelines: • There is a plausible temporary relationship between the onset of AE and the study drug and the AE cannot be easily Resolved or eliminated by the clinical condition of the subject, the intervening disease or accompanying therapy, and/or from a known formula that meets the response to the study drug, and/or after the study drug or dose is reduced, and (if Appropriate) Re-emerge after re-exciting. • There is no evidence that the AE has a pathogen other than the study drug (eg, pre-existing medical condition, underlying disease, intermittent disease or concomitant drug and/or AE does not have a plausible relationship with the study drug) (For example, the cancer annotation diagnosed 2 days after the first administration of the study drug: the researcher's assessment of the cause of the individual AE report is part of the research. With the individual eight £ report, ",, the deductor will immediately "Products of the product... (Port 5 evaluation does not matter) The product experience of the main W is evaluated in all cases and promptly communicates to the researcher and the appropriate regulatory agency that the observations may be sexually observed. The heart uses all the examiners to evaluate the 6-top method caused by the time point. Examples of non-directional inquiry include the following: & ° How does the feeling since the last clinical visit?,, since the first visit, there is any new Is the title?? Also < Health 126382.doc -67- 200826981 Specific instructions for recording adverse events on the CRF When recording AE or SAE on the CRF, the investigator should use Appropriate medical terminology/concept. Avoid proverbs and abbreviations. All AEs should be recorded on the AE crf page. There is a space on this page to indicate whether the event is serious (Y/N). For SAE, the SAE CRF page must also be completed. Record a medical concept in the event area on the SAE CRF page.

a. Relative to the diagnosis of signs and symptoms, if known at the time of reporting, the diagnosis should be recorded on the CRF instead of individual symptoms and symptoms (eg recording only liver failure or hepatitis instead of jaundice, flapping tremor and transaminase) The enzyme is elevated). However, if the cluster of symptoms and/or symptoms is not medically characterized as a single diagnosis or symptom at the time of reporting, the individual event § should be recorded as an AE CRF Page®. If a diagnosis is subsequently determined, it should be reported as tracking information. b. After other events, T ^ Generally, AEs (such as cascading events or clinical continuations) that occur after other events should be identified by the primary cause. For example, it is sufficient to record diarrhea as an AE CRF page only if it is known to cause severe laparotomy. However, if the major secondary AE is separated from the initiation event in time, both should be recorded as independent events. For example, if right severe bowel hemorrhage causes kidney failure, the event should be recorded as an independent AE CRF page. c• Persistent or recurrent adverse events The AE is the subject's assessment time. The interval between the ten points is extended and cannot be eliminated. 126382.doc •68- 200826981 In addition to the AE. Unless the severity increases, such events should only be recorded once in the crf. If the right persistent metamorphosis is more serious, it should be recorded again on the ae CRF page. Recurrent sputum is alpha ε that occurs, is eliminated, and subsequently recurs. All recurrent sputum should be recorded on the AE CRF page. d·Clinical laboratory abnormalities

In general, @individual laboratory anomalies are not recorded as AEs on the CRF. Clinically significant laboratory abnormalities that only cause study withdrawal, meet severity criteria, are related to clinical conditions or symptoms, or require medical intervention (eg, low hemoglobin requiring blood transfusion) are recorded on the AE CRF page. If a clinically significant laboratory abnormality is a symptom of a disease or syndrome (eg, alkaline phosphatase and bilirubin 5 times the upper limit of normal values associated with cholecystitis), then only a diagnosis (eg, cholecystitis) should be recorded in the AE (: On page 17. If a clinically significant laboratory abnormality is not a symptom of a disease or syndrome, the abnormality itself should be recorded on the AE CRF page. 1 The laboratory abnormality can be communicated as a clinical diagnosis. The diagnosis should be recorded as AE or SAE. The high serum potassium content of 7.0 mEq/L should be recorded as "hyperpotent potassium," and observations of clinically significant laboratory abnormalities should not be repeated on the AE CRF page, unless it is severe. , severity or pathogenic changes e. Pre-existing medical conditions first month] The i-pathology present is the medical condition present at the beginning of the study. These conditions should be recorded on the CRF page of medical and surgical history. The entire trial _ should be reassessed for pre-existing medical conditions and only recorded as ae = "AECRFW on the AECRFW surface when the frequency, severity or characteristic deterioration of the disease 126382.doc -69- 200826981 = At times, the mrr word conveys the concept of the pre-existing money 6 change (eg, headaches are more frequent). L death is not related to attribution, all deaths that occur during the AE reporting period of the program will be recorded on the AECRF page and quickly Report to the sponsor. When recording the event, the event or condition of the report should be recorded as a single-medical concept on the CRF page. ^ If the cause of death is unknown and cannot be determined at the time of reporting, record on the AE CRF page. "Unclear death". You should ride a variety of trials (for example, through a primary care physician, an autopsy report, a hospital record) and quickly report the cause of death to the sponsor. g. Hospitalization for medical or surgical procedures Treatment, any ae that results in hospitalization or extended hospitalization should be recorded and reported as SAE. If the subject is hospitalized for medical or surgical procedures due to AE' then the event of the procedure, rather than the procedure itself, should be recorded as sae For example, if the subject lives 9 to experience coronary artery bypass hand _, the bypass will become a necessary heart disease recorded as SAE. Hospitalization will not be recorded as a sae on the CRF: • Hospitalization or extended hospitalization of a pre-existing conditional diagnosis or elective surgical procedure that has not deteriorated in strength or frequency and recorded on the CRF of medical and surgical history 126382.doc -70- 200826981 • Allows for hospitalization or extended hospitalization required for study efficacy measurements • Hospitalization of scheduled treatments for extended disease of the study or extended hospitalization example 2 Objectives of the study clarified in this example To test the efficacy and safety of tenepase in restoring the function of dysfunctional HD catheters without placebo control. Objectives The objectives of the study were as follows: • To evaluate the safety of tenepase in the treatment of subjects with dysfunctional HD catheters. • Evaluation of the efficacy of tenepase to improve BFR in dysfunctional HD catheters This is a phase III, open-label study to be conducted in approximately 60 centers in the United States and Canada. About 225, 6 years old, requiring HD and having an arterial pressure defined at -250 mmHg during the first 30 minutes of HD (or a mechanical criterion of maximum negative arterial pressure, no more than 250 mmHg) BFR < 300 mL/min Subjects with functional abnormalities HD catheters that are at least 25 mL/min lower than the prescribed BFR will be enrolled in the study. The study will consist of a visit to the HD treatment period for each subject and a follow-up visit. Subjects will receive up to three open-labeled tenepase treatments: one or two treatments as part of the initial treatment process and one additional treatment as part of the retreatment procedure (if indicated). At the first visit, the first treatment of tenecteplase was injected into each lumen of the HD catheter for all subjects. After 1 hour of indwelling time, the subject is experiencing HD or experiencing HD to the extent possible as prescribed 126382.doc -71 - 200826981. At the beginning of HD, BFR was measured every 30 minutes thereafter, 30 minutes before the end of HD and at the end of HD to assess catheter function and determine treatment outcome. At the end of the first visit HD, subjects with BFR < 300 mL/min will have a second treatment, and the instillation will last for a longer period of time until the second visit (up to 72 hours). Prolonged at the beginning of the second visit * The indwelled tenepase will be withdrawn from the catheter and the subject will experience HD or experience HD to the extent possible. BFR is measured at the beginning of HD, every 30 minutes thereafter, 30 minutes before the end of HD, and at the end of HD. The first visit or the second visit was successful (defined as BFR2300 mL/min at arterial pressure and 0/250 mmHg at the end of HD and 30 to -250 mmHg at the end of HD and increased from baseline BFR by 225 mL/min) and abnormal dysfunction of the catheter within 21 days of the first visit (BFR at 30 min before arterial pressure HD at -250 mmHg [or no more than 250 mmHg under the mechanism of maximum negative arterial pressure]] Subjects at 300 mL/min and at least 25 mL/min lower than the prescribed BFR will withdraw from the initial course of treatment and enter the retreatment process, during which the subject will receive another tenecteplase Treatment (at the first visit to re-treatment [RT]). After 1 hour of indwelling time, the subject experienced HD or experienced HD to the extent possible as prescribed. BFR is measured at the beginning of HD, every 30 minutes thereafter, at the end of the HD knot ‘ 30 minutes before the bundle and at the end of HD. All subjects will undergo a follow-up assessment of HD catheter function at each of the two visits after the final study drug exposure. The second visit from the start of treatment to the final study drug exposure completed the recording of adverse events for all subjects. All subjects will undergo an antibody test after 30-36 days of the first visit or after the study is terminated early. Outcome measures 126382.doc -72- 200826981 Safety outcome measures The measurement of primary female complete results is as follows: • For the subjects who did not receive extended indwelling tenapes at the first visit: 'from the preliminary study The incidence of the first-degree female completeness of the drug was reported as follows: the incidence of intracranial hemorrhagic events (intracranial hemorrhage, major hemorrhage, blood pine, thrombosis, catheter-related bloodstream infection, and catheter-related complications) :

• In the first visit, patients who did not receive extended indwelling tenapes were s' from the start of the f2 visit to the third visit to complete the adverse events (as listed above). The first visit to the subject who extended the indwelling tenectease and the infusion of L's extended indwelling tenepase to the secondary treatment to complete the adverse events of the standard (as listed above) Subjects in the retreatment process, the incidence of target adverse events (as listed above) from the infusion of RT tenepase to the third visit to RT x • For the first visit, no extended retention was accepted For the subjects of Nyproxase, the incidence of serious adverse events from the initial study drug administration to the second visit and the incidence of all adverse events • For the first visit, the extended retention of tenep was not accepted. For the subjects of the enzyme, the incidence of serious adverse events and the incidence of all adverse events from the start of the second visit to the third visit • For the first visit, the extended stay of tenecteplase For the examinee, 'self-prolonged placement of tenet The instillation of the enzyme to the 4th visit was completed 126382.doc -73- 200826981 The incidence of adverse events and the incidence of all adverse events • For the subjects entering the retreatment process, the instillation from RT tenepase The incidence of serious adverse events and the incidence of all adverse events at the 3rd visit to RT • The incidence of positive anti-Temperpase antibody test in the subjects with negative baseline test Efficacy Results Measurements Primary efficacy results The measurements are as follows: • The first-time treatment of BFR treatment success (as defined above; see “Study Design π”) The percentage of subjects with secondary efficacy results measured as follows: • For the first visit to treatment success (eg above) For the subjects defined, the catheter function was maintained at the 2nd and 3rd visits (defined as BFR23 00 under arterial pressure in the range of 0 to -250 mmHg at the beginning of the HD treatment period (within the first 30 minutes) Percentage of subjects with mL/min and 225 mL/min from baseline BFR • As measured by blood urea nitrogen (BUN) before treatment and after HD at the first visit, there was a 265% reduction in urea (URR) Percentage of subjects • For those who did not receive extended retention of tenectease at the first visit, such as the percentage of subjects with a URR of 265% assessed by BUN measurements before and after the second visit to the HD • At the end of the first visit to HD BFR changes from baseline • Percentage of subjects of the following types defined by changes in BFR from baseline at the end of the first visit to HD: &lt;0 mL/min, 0-24 126382.doc -74· 200826981 mL/min, 25_49 mL/min, 50-99 mL/min, 100-149 mL/min, and 2150 mL/min For the first visit to patients with extended tenapopeptidase treatment, Level efficacy results measurements also include the following: • Percentage of subjects who were successful in BFR treatment (as defined above) at the 2nd visit • As assessed by BUN measurements before and after the 2nd visit to HD, the same 65% Percentage of subjects with URR • Percentage of subjects with a URR of 265% as assessed by BUN measurements before and after the third visit to HD • For subjects who succeeded in the second visit, in the third and Percentage of subjects who maintained catheter function (as defined above) at the 4th visit • At the end of the second visit to HD, BF Changes in R from baseline • Percentage of subjects of the following types defined by changes in BFR from baseline at the end of the second visit to HD: &lt;0 mL/min, 0-24 mL/min, 25- 49 mL/min, 50-99 mL/min, 100-149 mL/min, and 2150 mL/min For the subjects who entered the retreatment process, the secondary efficacy results were also measured as follows: • The first visit at RT Percentage of subjects with successful BFR treatment (as defined above) • Percentage of subjects with a URR of 265% as assessed by BUN measurements before RT first visit and after HD • As in RT 2 Percentage of subjects with 265382.doc •75-200826981 URR assessed by BUN measurements before and after HD visits. • For subjects who were successfully treated on the first visit to RT, RT 2 and 3 Percentage of subjects who maintained catheter function (as defined above) at the second visit • BFR changes from baseline at the end of HD at the first visit to the HD • Defined by changes in BFR from baseline at the end of HD at the first visit to HD Percentage of subjects of each of the following types·· &lt;0 mL/min, 0-24 mL/min, 25-49 mL/min, 50_99 mL/min 100-149 mL / min and 2150 mL / min square tenecteplase security design approved for reducing the acute myocardial infarction (AMI) associated mortality. Adverse events associated with systemic use of 30-50 mg of tenepase in the treatment of AMI are fully described and consist primarily of bleeding complications including major bleeding events and intracranial hemorrhage. Another adverse event that can be associated with the use of a thrombolytic agent to treat a malfunctioning catheter is a catheter-related thrombotic embolic event. Based on the clinical experience of tenepase in the treatment of AMI and CATHFLO® ACTIVASE® (alteplase) in the treatment of dysfunctional central venous access catheters, it is expected that any potential bleeding or thrombotic events attributable to tenecteplase are likely to be treated 24 Appeared within hours. All adverse events were recorded from the start of the study treatment to the second visit after the final study drug exposure. Study Treatment As described above (see "Research Design"'), depending on the recovery of HD catheter function, the subject will receive up to three doses of open-labeled tenecteplase. At each dose, 2 mL will be administered to the subject. (2 mg) tenecteplase was injected into each lumen of its HD catheter 126382.doc -76- 200826981. Companion therapy and clinical specification from the first visit to the second visit after the final study drug exposure It is forbidden to use fibrinolytic agents (except for research drugs: ==: forest)... or one or two::: for use between heparin or for prevention.) From the first visit to the final study, after exposure to the second dose After the completion of the visit, taking _8 (Cropido hydrogen sulfate fi salt tester may not increase the dose. Subjects can be treated in the treatment of _ _ _ _ _ other drugs and standards for their silk administration. Safety Analysis #Error is summarized by the body system, high-level terms and preferred terms from the preliminary study: the target adverse event in the subject who did not receive the extended indwelling tenepase dose at the time of the first visit to the second visit Incidence rate. Lost data for analysis purposes, Subjects who discontinued the study for any reason that did not achieve BFR 300 mL/mni and increased from baseline BFR &gt;25 rainbow/discussion would be considered a treatment failure. The sample size was determined to be 225 and the sample size was sufficient to adequately Accuracy estimates the incidence of relatively common adverse events. During the interim analysis period, DMC will conduct periodic reviews of cumulative safety data. The abbreviations used in this study are related to the abbreviations described in the example above. 126382.doc -77- 200826981 The detailed study design is a Phase III, open-label study conducted at approximately 60 centers in the United States and Canada. About 225 216-year-old subjects requiring HD and dysfunctional HD catheters will be enrolled in the study. The examiner will be classified into three categories by baseline BFR: 0-199 mL/min, 200-274 mL/min, and 275_299 mL/min. Number of registered persons in the 0-99 mL/min and 275-299 mL/min categories. Subjects will be limited to a maximum of 1% of each subject. The study will consist of a visit to the HD treatment period for each subject and a follow-up visit. During the study period, the subject will receive up to three open markers. Napase During the initial treatment, the subject will receive one or two treatments, and the eligible subject who becomes functionally abnormal again within 21 days of the first visit will receive additional treatment as part of the retreatment process. Informed consent (and, if available, child informed consent), screening qualified subjects at screening visits based on study inclusion and exclusion criteria (see sections 4.1.2 and 4.1.3). Screening at the discretion of the investigator The first visit and the first visit can be combined. At the first visit, the treatment with tenectease at the beginning of HD (the first 30 minutes) at -250 mmHg arterial pressure (or under the maximum negative arterial pressure, A qualified subject with a BFR of <300 mL/min and a BFR of at least 25 mL/min lower than the prescribed BFR at over 250 mmHg). The BFR measurement obtained at the first visit to determine the eligibility of the study was the baseline BFR. For the subject, 2 mL (2 mg) of tenecteplase was injected into each of the two lumens of the HD catheter. After 1 hour of indwelling time, the study drug was withdrawn and all subjects experienced HD or experienced HD as prescribed to 126382.doc -78 - 200826981 degrees. At the beginning of HD, every 30 minutes thereafter, BFR was measured 30 minutes before the end of HD and at the end of HD to assess catheter function and determine the outcome of the first visit (see above). As part of the initial treatment procedure, 2 mL (2 mg) of tenecteplase was injected into each lumen of the catheter for subjects with BFR &lt; 300 mL/min at the end of the first visit to HD. The treatment is left in an extended period of time until the second HD treatment period of the second visit (up to 72 hours). For those who received extended indwelling tenepase at the beginning of the second visit, the treatment was withdrawn from the catheter. Subjects experience HD or experience HD to the extent possible as prescribed. At the beginning of HD, every 30 minutes thereafter, BFR was measured 30 minutes before the end of HD and at the end of HD to assess catheter function and determine the outcome of the second visit. Subjects who have successfully treated at the first visit or the second visit and who have a recurrent dysfunction of the catheter during the first visit within 21 days (see Section 3. 1.2.b below) will withdraw from the initial treatment and enter the retreatment process. In this process, the eligibility is screened based on the retreatment inclusion and exclusion criteria (see Sections 4.1.4 and 4.1.5) and 2 mL (2 mg) of tenecteplase is injected into each lumen. Then stay for 1 hour (when re-treatment [RT] first visit). The BFR measurement used to determine re-treatment eligibility will be considered as the RT baseline BFR. At the beginning of HD, every 30 minutes thereafter, BFR was measured 30 minutes before the end of HD and at the end of HD to assess catheter function and determine the treatment outcome of the first visit to RT. A follow-up assessment of HD catheter function was performed by measuring BFR at the beginning of each HD (within the first 30 minutes) after two visits to the final study drug exposure. For these assessments, the researchers will increase BFR to attempt to achieve a prescription BFR of 126382.doc -79-200826981 within the first 30 minutes. If the HD catheter is removed at any time for any reason, no treatment is provided and no efficacy assessment (ie, BFR measurement or blood urea nitrogen [BUN] analysis) is performed. However, subjects will continue to undergo a safety assessment (ie, recording adverse events and accompanying drug and antibody tests). Subjects with symptomatic hypotension may not receive study medication. The second visit from the start of treatment to the final exposure of the study drug was recorded to record adverse events for all subjects. All subjects will undergo an antibody test after 30-36 days of the first visit or after the study has terminated prematurely. BFR Evaluation All BFR measurements in this study will be performed at arterial pressures ranging from 0 to -250 mmHg. During each HD treatment period, BFR should be increased until a local BFR is achieved. However, BFR should not be increased to the point where the corresponding arterial pressure exceeds -250 mmHg (eg -260 mmHg). At the 1-4th visit, each subject must be dialysis on the same type and model of HD device to maintain consistency of data collected during HD. For subjects entering the retreatment procedure, the same type and model of device must be used for the first 1-3 visits to the RT; however, the device may differ from the device used by the subject during the initial treatment. To be eligible for this study, subjects must have &lt;300 mL/min at an arterial pressure of -250 mmHg (or no more than 250 mmHg under the maximum negative arterial pressure criteria) during the first 30 minutes of HD. Baseline BFR at least 25 mL/min lower than the prescribed BFR. Subjects who are unable to measure BFR due to total obstruction (ie, no blood draw function) should be considered to have a BFR of 0 mL/min. 126382.doc -80 - 200826981 HD catheter line reversal During the study, all BFR data must be collected using a catheter line in the usual direction, ie the arterial (red exit) line of the HD catheter is used for blood removal and veins (blue The inlet) line is used for blood return. If the investigator determines that a reversal line is necessary to achieve fluid removal or electrolyte control, the BFR measurement must be recorded prior to reversal. Determination of treatment outcome, maintenance of catheter function, and recurrent catheter dysfunction The treatment success of this study will be defined as follows: at arterial pressure in the range of 0 to -250 mmHg, at the end of HD and 30 (10) minutes before the end of HD, BFR &gt; 300 mL/min and an increase of 225 mL/min from baseline BFR (wireless reversal). Subjects with BFR 2300 mL/min and increased from baseline BFR &lt; 25 mL/min, subjects with BFR &lt; 300 mL/min and catheter line reversal will be considered treatment failure. If the investigator determines that the subject's hemodynamics has become unstable (blood pressure drop or heart rate change) and therefore needs to reduce its BFR, then the BFR measurement must be recorded before the BFR is lowered. BFR within 30 minutes prior to hemodynamic instability will be used to determine treatment outcome. For subjects who were successful in the first visit or the second visit, maintaining catheter function at follow-up visits was defined as arterial pressure in the range of 〇 to -250 mmHg at the beginning of the HD treatment period (top 30 Within minutes) BFR 2300 mL/min and an increase of 225 mL/min from baseline BFR (wireless reversal). For these assessments, the investigator will increase the BFR in an attempt to reach a prescription BFR within the first 30 minutes. Recurrent catheter dysfunction was defined as arterial pressure BFR &lt; 300 mL/min at -250 mmHg during 30 minutes prior to HD (or no more than 250 126382.doc -81 - 200826981 mmHg under institutional guidelines for maximum negative arterial pressure) And at least 25 mL/min lower than the prescription BFR (use the catheter line in the usual direction). This open-label clinical trial was designed to assess the safety and efficacy of tenepopzyme in restoring the function of an abnormal HD catheter. The basic principles of the selected dose of tenecteplase are based on the approved dose of Cathflo Activase and the experience of using alteplase in CVA and HD catheters reported in the literature. Cathflo Activase is currently approved for the treatment of dysfunctional CVA devices. In clinical trials, up to two 2 mg doses of alteplase (body weight &lt; 30 kg subject dose lower) (up to 120 minutes indwelling time after each dose) effectively and safely restore dysfunctional CVA devices Features. There was no systemic administration in this study. However, in the case where the lumen size of the catheter is unknown or less than the prescribed dose of tenecteplase (2 mL) in this study, the possibility of one part of the administered dose (ie, the difference between the volume of 2 mL and the lumen of the catheter) enters the systemic circulation. Sexuality exists. The dosing regimen was not significantly different from the dosing regimen of Cathflo Activase clinical trials (A2055g, A2065g, and A2404g) in which a young subject weighing <30 kg was administered a dose equal to 110% of the lumen volume of the catheter. In the event that all 2 mg doses are not intended to be administered as an intravenous bolus injection, this would result in an expected maximum plasma concentration of 〇25 pg/mL. To best illustrate this, the maximum predicted concentration of 2 mg alteplase is 0.5 8 pg/mL. In comparison, the 30 mg dose of tenepase commonly used in AMI will result in a maximum plasma concentration in the range of 5.9 to 7.5 pg/mL (average data from the TIMI 10A and 10B trials) (Cannon et al. Circulation 1998). 98:2805-141 Cannon et al., Circulation 1997; 95:351-356). Similarly, Tanswell et al. (J Am Coll 126382.doc -82- 200826981

Cardiol. April 1992; 19(5): 1071-5) predicted that a maximum concentration of approximately 4 pg/mL could be achieved in patients given 100 mg of alteplase via an accelerated infusion regimen. In comparison, endogenously produced tissue plasminogen activator levels have been reported to be in the range of 0.002 to 0.021 pg/mL. The basic principle of limiting the BFR &lt; 300 mL/min to the arterial pressure of -25 0 mmHg is based on the recommendation in the KD0QI guidelines for vascular channels of hd, which requires 23〇〇mL/mintBFR Provide adequate dialysis without limiting the duration of the HD treatment period. Since BFR is directly related to negative arterial pressure, this study set the arterial pressure to _25〇 mmHgW to maintain consistent conditions for BFR determination. In addition, the &amp;£&gt;〇卩1 guidelines recommend measuring BFR at an arterial pressure of -250 mmHg to determine catheter dysfunction. Safety outcome measures Primary safety outcome measures are as follows: • For subjects who did not receive extended indwelling tenapes at the first visit, the initial dose from the initial study drug to the second visit The incidence of secondary safety results for adverse events, major bleeding, sputum events, blood tests, catheter-related bloodstream infections [CRBSI], and catheter-related complications were as follows: W enzymes were examined Target adverse events for the first time in the first visit without receiving extended indwelling tenapril, from the start of the second visit to the completion of the third visit (as listed above) for the extension of the first visit Indwelling

126382.doc -83· 200826981 • For the subjects who entered the retreatment process, the incidence of adverse events (such as those listed above) from the infusion of RT tenepase to the third visit to RT • For those who did not receive extended indwelling tenepase at the first visit, the incidence of serious adverse events from the start of the second visit to the second visit and the incidence of all adverse events • For those who did not receive extended indwelling tenepase at the first visit, the incidence of serious adverse events from the start of the second visit to the third visit and the incidence of all adverse events • The incidence of serious adverse events and the incidence of all adverse events from the infusion of extended indwelling tenepase to the fourth visit for the first time patients who received extended indwelling tenecase at the first visit • For the subjects who entered the (four) course of treatment, the incidence of serious adverse events from the infusion of RT tenepase to the third visit to RT and all adverse events occurred • The test was negative at baseline Positive anti-tenapase resistance Results The incidence of efficacy of the test results of the amount of measurement as the primary efficacy measure in the first - ·· people do BFR percent on treatment success (as defined above) of the subject. The first-level efficacy results are measured as follows: · For the subjects who successfully treated the first visit, the percentage of subjects who were in the second and third visits for the #catheter function (as defined above) was 126382.doc - 84- 200826981 • Percentage of subjects with a 265% reduction in urea (URR) as assessed by BUN measurements at the first visit at the first visit • For those who did not receive extended tenapes at the first visit For the examinee, the percentage of subjects with a URR of >65% evaluated by the BUN measurement before and after the second visit to the HD: • The change in BFR from the baseline at the end of the first visit to the HD • By the first Percentage of subjects of each of the following types defined by changes in BFR from baseline at the end of HD visit: &lt;0 mL/min, 0-24 mL/min, 25-49 mL/min, 50-99 mL/ Min, 100-149 mL/min, and ^150 mL/min For subjects who were treated with extended tenapectase at the first visit, the secondary efficacy results were also measured as follows: • At the 2nd time Percentage of subjects who were treated successfully for BFR treatment (as defined above) • As assessed by BUN measurements before and after the second visit to HD &gt; Percentage of subjects with a URR of 65% • Percentage of subjects with a URR of 265% as assessed by BUN measurements before and after the third visit to HD • For those who succeeded in the second visit Percentage of subjects who maintained catheter function at the 3rd and 4th visits (as defined above) • BFR changes from baseline at the end of the second visit to the HD • BFR from the end of the second visit to HD The change in baseline defines the percentage of subjects of each of the following types: · 0 mL/min, 0-24 mL/min, 25-49 mL/min, 50-99 mL/min, 100-149 126382. Doc -85- 200826981 mL/min and >150 mL/min For subjects entering the retreatment process, the secondary efficacy outcome measures also include the following: • The first visit to the BFR treatment at RT (eg above) Percentage of subjects in the definition) • Percentage of subjects with a URR of 265% as assessed by BUN measurements before and after the first visit to RT • For example, the BUN measurement before and after HD at the second visit to RT Assess the percentage of subjects with a URR of 265% • For subjects who were successfully treated on the first visit to RT, at RTs 2 and 3 Percentage of subjects who maintained catheter function (as defined above) • BFR changes from baseline at the end of HD at the first visit to the HD • Belong to the definition of BFR from baseline at the end of HD at the first visit to HD Percentage of subjects of each type: &lt;〇mL/min, 0-24 mL/min, 25-49 mL/min, 50-99 mL/min, 100-149 mL/min, and &gt;150 mL/ Min Safety Design Tenepase is approved for reducing mortality associated with AMI. Adverse events associated with systemic use of 30-50 mg of tenepase in the treatment of AMI are fully described and consist primarily of hemorrhagic events including major bleeding events and ICH. The elimination of tenecteplase from plasma is divided into two phases with an average initial half-life of 20-24 minutes and an average final half-life of 90-130 minutes (Modi et al. above). Although the incidence of bleeding complications in subjects with AMI treated with tenepase was quantified, the bleeding associated with the lower 126382.doc -86-200826981 dose of tenepase used in this study There is limited information on the incidence of complications. The incidence of ICH and major bleeding due to tenecteplase is expected to be relatively low in this study due to the proposed low dose, minimal systemic exposure to tenecteplase and the use of CATHFLO® ACTIVASE® (alteplase) to date. Clinical trial experience indicating that ICH was not reported and only 3 of the 1432 subjects experienced major bleeding. Another adverse event associated with the use of thrombolytic agents to treat dysfunctional catheters is catheter-related thrombotic embolic events. This event can cause pulmonary embolism, which can be life-threatening depending on the size of the pulmonary embolism. The extensive experience of using urokinase and Cathflo Activase in CVA catheters is expected to be associated with a lower incidence of clinically significant seizure events associated with the use of tenectease clearance catheters. Based on the clinical experience of tenaplase in the treatment of dysfunction of AMI and Cathflo Activase CVA catheters, it is expected that any potential bleeding or thrombotic events attributable to tenecteplase will likely occur within 24 hours of treatment. All adverse events were recorded from the start of the study treatment to the second visit after the final study drug exposure. All serious adverse events will be reported to Genentech within 48 hours, regardless of cause or treatment. See Section 5 for full details of the safety assessment for this study. The Data Monitoring Committee (DMC) DMC will review cumulative safety data for the tenectease catheter removal program at scheduled intervals, including dysfunctional CVA and HD catheters (study of N3 698g, N3 699g, N3 700g, and N3 701g) And based on the results of the review process, it is responsible for making recommendations to the sponsor regarding the continued safety of the research. 126382.doc •87- 200826981 Study Subjects and Analysis Group Subjects with functionally abnormal HD catheters are eligible for this study and will be screened using the criteria provided herein. There was no control group in this study. Compliance with laws and regulations This study is conducted in accordance with the requirements of the U.S. FDA, INTERNATIONAL CONFERENCE ON HARMONISATION E6 GUIDELINE FOR GOOD CLINICAL PRACTICE (GCP) and any country. Materials and Methods Subject Selection for Subjects Subjects with a functionally abnormal HD catheter (as defined above) based on BFR during the first 30 minutes of HD were eligible for this study. Approximately 225 subjects from approximately 60 research sites in the United States and Canada will be enrolled. Subjects will be classified into three categories by baseline BFR: 0-199 mL/min, 200-274 mL/min, and 275-299 mL/min. The number of registered persons in the 0-199 mL/min and 275-299 mL/min categories will be limited to a maximum of 1% of the subjects in each category. Use the inclusion and exclusion criteria listed below to screen subjects. Subjects included in the standard must meet all of the following criteria to be eligible for inclusion in the study: • Can provide written informed consent and follow the study evaluation for the entire duration of the study • Age 216 • Researcher believes clinically stable • Use Sleeve tunnel type HD catheter with a BFR of &lt;300 mL/min at a maximum negative artery of 126382.doc -88· 200826981 at 250 mmHg, but BFR at least one HD treatment period 7 days before the first visit Prove 2300 mL/min • HD BFR 2300 mL/min • Under arterial pressure of -250 mmHg (or no more than 250 mmHg under the mechanism of maximum negative arterial pressure) &lt; 300 mL/min baseline BFR (HD During the first 30 minutes) (use the catheter line in the usual direction before any reversal of the line; see section 3 · 1.2.a) • Baseline BFR (during 30 minutes before HD) is at least 25 mL/min lower than the prescription BFR eg HD Subjects with a prescription BFR of 300 mL/min must have a BFR of £275 mL/min to enter the study. • At an arterial pressure ranging from 0 to -250 mmHg in at least one HD treatment period 7 days prior to the first visit, BFR proved to be >300 mL/min (using a catheter line in the usual direction) • Expect at least 30 The same catheter is used on HD devices of the same type and model. • Exclusion criteria for volume infusion fluid studies necessary for injecting study drug into HD catheters. Subjects that meet any of the following criteria will be excluded from the study. : • HD catheter with a BFR of 2300 mL/min after the subject was reset • HD catheter insertion before screening • 2 days • Mechanical, non-thrombotic causes of abnormal dysfunction of the HD catheter (eg catheter writhing or compression catheter) Sewing) or evidence of dysfunction caused by known fibrin sheaths 126382.doc -89 - 200826981 The use of implantable 埠HD catheters not implanted in the subclavian vein is expected to be used during the study for any other Type of procedure for the conduit (ie not for HD)

Previously treated or any tenectease catheter clearance test using fibrinolytic agents (eg, enzyme, tenecteplase, ribine) within 7 days of the first visit using any study drug or therapy within 28 days of: k刖普Enzyme or urokinase) HDWhen the screening is known to prematurely or secrete milk with a known or suspected infection of the HD catheter • History of any intracranial hemorrhage, aneurysm or arteriovenous malformation • Use any within 24 hours before the first visit Heparin (ordinary or low molecular weight), except for heparin used only during HD or for prevention (eg heparin lock) μ • Warfarin is used within 7 days prior to the first visit, except for low-dose warfares for prevention Lin Wai • Start or increase the dose of Plavix® (croprozil bisulfate) within 7 days before the first visit

• In the case of red jk pheromone, hemoglobin &gt; 1 3.5 g/dL Laboratory test for hemoglobin content must be performed within 3 days prior to screening. • Platelet count &lt; 75,000/μί Laboratory test for confirming platelet count Must be performed within 3 days prior to screening 0 126382.doc -90- 200826981 • The investigator believes that a bleeding event or embolic disorder (ie, recent pulmonary embolism, deep vein thrombosis, endarterectomy, or clinically significant right to left High risk of shunting or known symptoms with significant risk of bleeding • BFR &lt; 300 mL/min due to symptomatic hypotension. • Researchers believe uncontrolled hypertension (eg systolic blood pressure > 185 mmHg and Shu - Tensile &gt; 110 mmHg) • It is known that there is a hypersensitivity to any component of tenecteplase or a formulation. The standard subject must meet all of the following criteria to be suitable for retreatment included in the study. In the process: • Successful treatment at the first visit or the second visit (as defined above) • The investigator believes that it is clinically stable • Continue to use the same sleeve type, tunnel type HD catheter (ie The catheter of the subject is in place during the initial treatment) • HD BFR 2300 mL/min I • Under arterial pressure of -250 mmHg (or no more than 250 mmHg under the mechanism of maximum negative arterial pressure) &lt; RT baseline BFR of 300 mL/min (use the catheter line in the usual direction before any reversal of the line; see section 3.1.2.a)

• RT baseline BFR at least 25 mL/min lower than the prescribed BFR For example, a subject with a BFR of 300 mL/min in HD must have a BFR of $275 mL/min to enter the retreatment procedure. • It is expected that at least three consecutive HD treatment periods will use the same catheter on the same type and model of HD device 126382.doc -91 - 200826981 can be met by the exclusion criteria for the volume of infusion fluid re-treatment required to inject the study drug into the hd catheter Subjects of any of the following criteria will be excluded from the study. · "No catheter with a constant of 0 / / mL after the subject is reset" · Mechanical, non-thrombogenic causes of abnormal HD guide function (eg catheters. kinks or compression catheter sutures) or evidence of dysfunction induced by known fibrin: • catheters intended for use in any other type of diagnostic or therapeutic procedure during retreatment of the study (also That is not for hd) &quot; • Use any research drug or therapy other than tenectease within 21 days prior to the first visit to the RT • RT use in addition to tenectease in the study within 7 days prior to the first visit Fibrinolytic agents (eg alteplase, reteplase, or urokinase) • Known for pregnancy or lactation during a visit to an RT brother 1 • HD catheter with known or suspected infection • Any cranial History of bleeding, aneurysm, or arteriovenous malformations • Use any heparin (ordinary or low molecular 1) within 24 hours prior to the first visit to RT, except for heparin used only during HD or for prevention (eg heparin lock) μ • RT uses warfarin within 7 days prior to the first visit, except for low-dose warfarin for prevention • RT starts or increases the dose of clopidogrel in the first 7 days prior to the first visit. In the case of hemagglutinin, the initial treatment process is known with rt 1 126382.doc -92- 200826981

Hemoglobin between the visits 213 5 g/dL • The platelet count between the initial treatment and the RT visit is known to &lt;75?000/μΕ • The investigator believes that ash events or embolic complications (ie, recent High risk of pulmonary embolism, bead vein blood test, endarterectomy or clinically significant right-to-left shunt) or known symptoms with significant risk of bleeding • BFR&lt;3〇〇mL/ due to hypotensive symptoms Min • Researchers believe that uncontrolled hypertension (eg systolic blood pressure &gt; 185 mmHg and diastolic blood pressure &gt; 110 mmHg) • It is known that there is a hypersensitivity formulation for any component of tenecteplase or formulation to have DAIKY The disposable, 6 cc glass vial of the 〇TM stopper and the easy-to-open aluminum cover provides tenecteplase. Provides nepeptidase··1〇4·4 mg arginine, 32 mg phosphoric acid and 〇·8 mg polysorbate 2〇 as a sterile, dry formulation containing 2 mg of protein with the following excipient specifications . The diluent used was USP/EP (;SWFI;) for injection sterilization. Dosage, Dosing, and Storage Depending on the recovery of HD catheter function, the subject will receive up to 3 times of tenapes. At each treatment, 2 mL (2 mg) of nepeptase was injected into each lumen of the HD catheter for the subject. If the HD catheter is removed at any time for any reason, no further treatment is provided. Subjects with symptomatic hypotension may not receive study medication. Each vial of lyophilized tenaplase was reconstituted immediately prior to use with 2·2 mL of BWFI. Use sterile techniques to direct BWFI directly into the study drug 126382.doc -93- 200826981 freeze-dried block and gently vortex the vial until the contents dissolve. Not shocking. The concentration of tenecteplase in the resulting solution was 1 mg/mL. Slightly foaming after rehydration is not anomalous; any large bubbles will disperse if the vial is allowed to sit quietly for a few minutes. If the reconstituted research drug is not used immediately, the solution must be stored at 2 -8 ° C (36 ° F - 46 ° F) and used within 8 hours of rehydration. Discard any unused solution.

Any fluid in the lumen of the HD catheter was withdrawn and attempted to flush with saline only prior to administration of the study drug. To dose, sterile 2 mL of the reconstituted study drug should be pumped into a single 1 mL syringe. According to the agency's guidelines, the solution should then be dripped into the lumen of a HD catheter. The remaining volume of the catheter should be filled with physiological saline. The second cavity is repeated. The vial. Do not use more than 2t-8t: (36T-46T) frozen storage of research drugs Store any unused parts for future use of vials.

Genentech provides a study drug on the vial or expiration date on the expiration date. Partially used vials, empty vials and unrehydrated vials will be returned to Genentech 剂量 Dosage changes No dose changes are allowed. Accompanied and excluded therapy, ^ Huan will not be allowed to receive any intravenous therapy or in the study drug is in ^ ^, work by the HD ^ official blood sample. Only by using an independent route, the intravenous sputum method or the medium 4 can be used to deliver blood samples. From the first visit to the second time after the final study drug exposure, the sputum is smashed into more than one fiber. Other than protein soluble research drugs), Huafa Yao, Hua Tinglin (except for low-dose warfarin for prevention) and 126382.doc -94- 200826981 Ordinary or low molecular weight heparin (except for use only during HD or for Prevention of heparin). The second visit from the first visit to the final study drug exposure is performed by the subject who took Cropirone Hydrochloride without increasing the dose. If you are under the treatment, you can continue to receive other drugs and standard treatments for your condition. Study Evaluation IX consists of a visit to the continuous He treatment period based on the regular HD time course of each subject and a visit. At the discretion of the investigator, 4 = combination of 5 townships and the first visit. Subjects will receive up to three times of nepase treatment. The first two treatments are part of the initial treatment process and one is &amp; treatment is part of the retreatment process. At the ith visit, the first home treatment == subject is followed by one hour of indwelling time. In the first...: diagnosis, the duration of the indwelling time is extended to the qualified subject for the second treatment until the second visit (up to 72 hours). At the first visit, or if the person is treated successfully and the dysfunction in the 21st day of the first visit (such as the female unit 6 ^ ^, 疋 meaning), the subject will withdraw from the initial treatment process ^ re-treatment process, in In this process, the eligible subject will receive another agent, two right enzymes, followed by a one-hour indwelling time (every time after re-treatment for the first time, the examiner will receive two visits after the final study drug exposure: experience Tracking evaluation of TM catheter function. According to different research drugs. The first visit is 30 (up to mm U and then the examiner will return to pursue the visit. After the study or early termination of the study, all at any time is out of any Reasons for removal of the HD catheter will no longer provide treatment 126382.doc •95- 200826981 and will no longer be evaluated for efficacy (ie BFR or BUN analysis). However, the examiner will continue to undergo a safety assessment (also Record adverse events and accompanying drug and antibody tests.) Hunting is provided by the central laboratory Quintiles Laboratories (QLab) to provide a laboratory kit and instructions for collecting Bun and anti-teinase antibody samples. All samples will be in situ. Processed and shipped to QLa b. QLab will perform a bun analysis, calculate and ship the antibody samples to Genentech for testing. Screening visits At the discretion of the investigator, any or all screening assessments can be performed at the third visit (before registration). Written informed consent/consultation must be obtained prior to the procedure. The following screening assessments and procedures will be implemented: • Written informed consent/consultation review and exclusion criteria review (see Sections 41.2 and 413). Includes the date of birth, gender and racial/ethnic physical examination and medical history, including two recent URR values (historical baseline). If physical examination is not indicated at the screening visit, a physical examination may be used. The restriction is that it is performed within 7 days before screening. Signs of sputum, including blood pressure, respiration rate, temperature and pulse (detailed if before and after) • Weight (detailed before and after HD) • If before screening 30 Blood samples are taken to determine hemoglobin content if laboratory tests are not performed within the day to confirm eligibility (if the subject is for red blood cells) 126382.doc -96- 200826981 素) and platelet counts • Accompanying drugs • HD catheter history and information records Information on the date of HD catheterization and the date of the last known HD catheter function (BFRdOO mL/min). Hd catheter lumen size, type, volume, brand (if known) and placement. The first visit to the HD prescription for the first visit within 7 days (as described above, screening and treatment at the discretion of the investigator) One visit can be combined. At the beginning of the first visit, the following should be performed to confirm eligibility: • Review of study inclusion and exclusion criteria (see sections 4.12 and 4.13) • Accompanied by medications and medical history Review (to ensure no change from screening), including the use of fibrinolytic agents, warfarin and crotidol bisulfate within 7 days prior to the first visit and the use of heparin within 24 hours prior to the first visit (See Section 4.1.3) • HD Prescription • HD, prescribed by prescription • Baseline BFR, measured at the beginning of HD (within the first 30 minutes) to confirm HD catheter dysfunction at -25 0 mmHg arterial pressure (or At maximum negative Under the mechanical guidelines of the pressure, no more than 250 mmHg) subjects with &lt;300 mL/min and a BFR lower than the prescribed BFR (using a catheter line in the usual direction) of at least 25 mL/min are suitable for this study; all others The subject is not suitable. Baseline BFR was recorded at 126382.doc-97-200826981 once to determine that the subject was eligible for the study (i.e., when the arterial pressure reached -25 0 mmHg). If dialysis with a reverse line has been attempted (i.e., prior to selection of study medication), BFR must be recorded prior to online reversal and used as a baseline value. Subjects who are unable to measure BFR due to total obstruction (ie, no blood draw function) should be considered to have a baseline BFR of 0 mL/min. A qualified subject will interrupt their HD. Unqualified subjects may complete their HD treatment period and should be registered as a screening failure. After the eligibility has been confirmed, the subject is registered using the Interactive Voice Response System (IVRS). The following assessments and procedures were also performed at the first visit: • Blood samples collected for serum anti-Tenapase antibody test before treatment with tenecteplase • Collected for BUN analysis prior to treatment with tenecteplase Blood samples • Tennipase was administered as described in Section 4.3.2 and left to rest in the subject's HD catheter (two lumens) for 1 hour before returning to HD. After leaving for 1 hour, the tenecteplase was withdrawn. • Subjects with symptomatic hypotension may not receive study medication. • Recover and perform HD as planned and resume and perform HD to the extent possible • At the beginning of HD, every 30 minutes thereafter, measure BFR 30 minutes before the end of HD and end of HD to determine treatment outcome (eg 3.1.2. As defined in Section b) If it is necessary to reverse the catheter line dialysis, the BFR measurement is recorded prior to reversal and no other BFR measurements will be recorded during this HD treatment period. If the investigator determines that the hemodynamic instability, the subject needs to reduce the bfr of 126382.doc -98- 200826981, the BFR measurement will be recorded before the BFR is lowered. The follow-up will continue to be recorded as scheduled. • Collect blood samples for BUN analysis after completion of HD If the catheter line has been reversed, no blood samples will be taken. • Adverse events and changes in accompanying medications during this visit The study began to monitor adverse events after initiation.

Nipase treatment in HD knots study drug in subjects HD

Subjects with BFR &lt; 300 mL/min at the second replacement as described in Section 4.3.2. The catheter (2 lumens) was left in a quiet place until the second hd treatment period (up to 72 hours) at the second visit. Subjects with symptomatic hypotension may not receive study medication. The second visit was performed on the following 2 times and the following assessments and procedures were performed: • Adverse events after the last visit and changes in accompanying medications • For the first time, only the first visit to the extended indwelling tenapes, pre-HD catheter Removal of extended indwelling tenepase (drip == time) • HD prescription • Blood sample collected for BUN analysis before HD is discarded if the subject becomes suitable for retreatment based on the initial BFR (see below) ). Λ ' • HD, starting with prescription (four) 'measured at the beginning of HD (within the first 30 minutes) to assess HD catheter function for 5 hai, etc., the researchers will increase the attempt to try the first minute 126382.doc - Prescription BFR is achieved within 99-200826981. Successful treatment at the first visit but with recurrent catheter dysfunction (as defined by the scabbard) and suitable for retreatment at the beginning of the second visit will interrupt the HD, $ initial treatment and entry Re-treatment (more common 5.6 mouth P knife). All other subjects will continue their prescription treatment period to the extent that they can perform the following assessments and procedures: • Subjects who receive extended extended tenectease only at the first visit

And a • At the beginning of HD, every 30 minutes thereafter, the BFR is measured 30 minutes before the end of HD and the end of HD to determine the treatment result (such as the upper definition of 3丄2). If it is necessary to reverse the catheter line dialysis, then before the reversal The (4) measurements are recorded and no other (four) measurements will be recorded during this HD treatment period. If the investigator determines that the subject needs to reduce his or her BFR due to hemodynamic incompatibility, the SCAN will be recorded before BFR is reduced. Subsequent brewing will continue to be recorded as scheduled. • Collect blood samples analyzed with kBIjn after HD is completed. If the catheter line has been reversed, no blood samples will be taken. Adverse events and changes in accompanying medications during the third visit The following assessments and procedures were performed at the third visit: • Adverse events and accompanying medication changes after the last visit • HD Prescription: Pre-HD collection for Blood sample for BuN analysis 126382.doc -100 - 200826981 If the subject becomes suitable for retreatment based on the initial BFR, the dragon discards it (see below). • Perform HD or perform HD to the extent possible • BFR, measure at the beginning of HD (within the first 3 minutes) to evaluate rib function g such as 30 minutes for this #evaluation, the researchers will increase BFR in an attempt to The prescription BFR is reached within. Subjects who are eligible for retreatment at the first or third visit or have recurrent catheter dysfunction (as defined in section 3丄3.b) and who are eligible for retreatment at the start of the third visit will have their HD Interrupted, $ out of the initial treatment process and entered the retreatment process (see section 4.5.6). All other subjects will continue the HD treatment period and perform the following assessments and procedures. • For: Subjects who receive extended indwelling tenecase only at the ith visit: Collected for HD after completion of HD Blood samples analyzed If the catheter line has been reversed, no blood sample is taken. Adverse events and changes in accompanying medications during this visit. 4th visit Only the second visit to the patient who received the extended tenapopeptidase requested the 4th visit. The subjects will perform the following research assessments and procedures: • Adverse events and accompanying medication changes after the last visit • HD prescriptions • Perform HD or HD implementation to the extent possible • BFR, at the beginning of HD Measurements (within the first 30 minutes) to evaluate the ^^^ catheter function For these assessments, the study will increase (4) in an attempt to achieve a prescription BFR within the first 3 minutes ^ 126382.doc -101 - 200826981. Subjects who are eligible for retreatment at the first visit or the second visit but have recurrent catheter dysfunction (as defined in Section 4, Section 4.b) and who are eligible for retreatment at the beginning of the fourth visit will interrupt their HD , exit the initial treatment process and enter - retreatment process (see section 4.5.6). All other subjects will continue their HD treatment period and perform the following assessments and procedures: • Adverse events and accompanying medication changes during this visit (the first visit or the second visit is successful) Subjects with recurrent catheter dysfunction (as defined above) within 21 days of the first visit will withdraw from the initial treatment process and enter the retreatment process, based on the inclusion and exclusion criteria for retreatment (see above) Description) Screening for eligibility, and it is treated with f-tenectilase once, followed by a one-hour indwelling time. It is expected that HDs of the same type and model should be used for all two-person retreatment HDs. Description The re-treatment process will consist of three visits. C, the second, third or fourth visit (if appropriate) or all of the pre-entry treatments will omit all related to the initial treatment process. Continue to see the doctor and complete only three re-treatment (RT) visits. For example, if the i-th treatment is successful and the second visit begins, the patient with recurrent catheter dysfunction is found. The assessment and procedures for the second visit will then be carried out instead of the second visit. Therefore, in this study, the subject will perform the assessment of the first visit, the RH-3 visit and the 3 day follow-up after the i-th visit. Procedure: a· RT first visit 126382.doc -102- 200826981 Use 1VRS to register a subject suitable for retreatment. The following assessments and procedures are performed on the subject at the first RT visit: • Make the subject appropriate BFR for retreatment (RT baseline BFR) • Review of inclusion and exclusion criteria (see Sections 4.1.4 and 4.1.5) Continuous tracking of subjects who do not meet these criteria according to the time course outlined in the initial treatment process Γ • Updated accompanying medications and medical history • HD prescriptions • Blood samples collected for BUN analysis prior to treatment with tenepase • Pre-recovery of HD before administration of tenectilase as described in Section 4.3 · 2 Nipproxase is allowed to remain in the subject's HD catheter (two lumens) for a few hours. After ten minutes of indwelling, tenecteplase is withdrawn. Subjects with symptomatic hypotension may not receive the study drug. • Recover and execute HD or restore and execute HD as possible • At the beginning of HD, every 3 minutes, 3 weeks before the end of HD and 4 minutes before the end of HD, measure BFR to determine the treatment outcome (as defined in Section 3.12b). If it is necessary to reverse the catheter line dialysis, then reverse The bfr measurement is recorded before and other b ρr measurements will not be recorded during this HD. If the investigator determines that the subject needs to lower his BFR due to hemodynamic instability, the BFR will be recorded before the BFR is lowered. Measurements • Blood samples collected for BUN analysis after completion of HD will not be collected if the catheter line has been reversed. This is a rare event and adverse drug changes during the period 126382.doc -103 - 200826981 b. RT 2 The following assessments and procedures will be carried out at the 2nd and 3rd visits to the RT. • Unfavorable events after the last visit and changes in accompanying medications • HD prescriptions • Only the second visit to RT·· Blood samples collected for BUN analysis before HD • Perform HD or HD implementation to the extent possible 'BFR at the beginning of HD (within 3 〇 minutes) to assess rib catheter function for these assessments, study Personnel will increase to try to be within the first minute A prescription BFR is reached. • RT only 2nd visit: blood samples collected for BUN analysis after HD completion If the catheter line has been reversed, no blood samples will be taken. The adverse events and accompanying medication changes during the visit were tracked on the 30th day or early termination. On the 30th day after the first visit (up to 36 days) or after the study was terminated, all subjects were taken. 9 anti-Tenapase antibody test. Information about the status of the catheter was also collected at the clinic. If an adverse event occurs before the second visit after the last study treatment, the adverse events are recorded at the early termination. Subjects are stopped The subject has the right to withdraw from the study at any time. The investigator has the right to withdraw the subject for any reason in the best interests of the subject, including intersequent illness, adverse events, or worsening of the condition due to a violation of the program, management reasons, for any reason, or To terminate the study's decision or any other valid and ethical reasons, Genentech reserves the right to request the subject to withdraw from 126382.doc -104- 200826981. Research stop

Genentech reserves the right to terminate the study at any time. Reasons for termination of the study may include, but are not limited to, the following: X or “The incidence or severity of adverse conditions in his study indicates a potential health hazard to the subject. • The registration of the subject is unsatisfactory. Data records are inaccurate or incomplete. Statistical Methods, ☆ This is an open-label one-arm study. All subjects enrolled and treated with tenecteplase will be included in the safety and efficacy analysis. The study summarizes the number of registered people and the reasons for the stop and stop of the dose of Tenepase. The safety analysis uses the MedDRA Dictionary to translate the literal description of emergency adverse events into better terms and body system terminology. The results were measured from the time when the initial study drug was administered to the second time = the initial rate of the recipient who did not receive the extended indwelling tenepase dose was less than one. The target adverse event was summarized in the target adverse event category. : The occurrence of all adverse events and serious adverse events at this same interval: a similar overview of the f::, the South Standard term and the preferred term. Generated from the 2nd 訧β A similar overview of target adverse events, all adverse events, and serious adverse events from the end of the third visit. 126382.doc -105- 200826981 : For subjects receiving extended indwelling doses of tenecteplase The adverse events category will outline the self-injection and extension of the indwelling dose to the fourth occurrence of the medical treatment. The same interval will result in the system and system of all adverse events and serious adverse events of the subjects. A high-level terminology similar to that of the father's second language. It is expected to be extended by the retention of tenecteplase, and will be produced from the initial tenectease to the extension of the indwelling tenapplase. An overview of adverse events, serious adverse events, and all adverse events.

C:) ^ In the case of recurrent treatment of recurrent catheter function with different t(tetra) nepase, Lu, in the category of no adverse events, the self-administered RT tenecteplase to RT at the end of the third visit Adverse events. A similar overview of the physical systems, high-level terms, and preferred terms for all adverse events and serious adverse events of these patients at the same interval. Efficacy analysis a. Primary efficacy outcome measurement The primary efficacy outcome measure is the percentage of subjects who were successful in the i-th treatment for coffee treatment (as defined above). It is considered that the study is stopped before the completion of HD or the primary outcome measurement is not evaluated. The subject is not satisfied with the primary outcome measurement. Calculate the percentage of subjects who achieved treatment success and provide a 95% confidence interval based on an accurate method. Sensitivity analysis was performed to assess the robustness of the primary outcome of the surrogate missing data approach, including complete case analysis and last observation advancement (LOCF) estimates. b. Secondary efficacy outcome measurement For subjects who have successfully completed the first visit, the catheter function will be maintained at the 2nd 126382.doc -106-200826981 and 3rd visit (as defined above) The percentage of subjects was analyzed similar to the measurement of primary efficacy results. Calculate the percentage of subjects who maintain catheter function at each visit and provide a 95% confidence interval based on an accurate method. Successful treatment at the first visit, discontinuation of the study before the completion of the 2nd and 3rd visits, or a BFR unpredictable at the time of such visit will be considered as a failure to measure the secondary outcome measurement. At the first visit and the first visit to the RT, the URR is calculated as follows: (BUN before treatment) - (BUN after HD) (BUN before treatment) At all other visits, the URR is calculated as follows: (pre-HD BUN)- (BUN after HD) (BUN before HD) At the 1-3th visit and the 1st and 2nd visits to RT, the percentage of subjects with a URR of 265% was calculated and a 95% exact confidence interval was provided. Subjects who prematurely discontinue the study or URR non-valuable will be considered to have a &lt;65% URR for this result. Summarize the mean change in BFR from baseline at the end of the first visit to HD and provide a 95% accurate confidence interval. The following change categories will also be used to analyze BFR changes from baseline at the end of the first visit to HD: &lt;0 mL/min, 0-24 mL/min, 25-49 mL/min, 50-99 mL/min, 100 -149 mL/min and >1 50 mL/min. For these results, subjects who missed BFR data will use the LOCF method to estimate the value. Use the alternative estimation method to assess the robustness of the results. Results of retreatment with regard to administration of extended indwelling tenecteplase and tenecteplase 126382.doc -107- 200826981 Measurements were performed similar to the above primary and secondary results measurements. C. Subgroup analysis provides estimates of the primary and secondary efficacy outcome measures and confidence intervals and key safety outcomes for the following subgroups: • Age: &lt;18, 18-65, &gt; 65 years • Gender: Male • Female • Baseline BFR: 0-199 mL/min, 200-247 mL/min, 275-299 mL/min Loss data for analysis purposes, discontinuation of study for any reason, failure to achieve treatment (as defined herein) The subject will be considered a treatment failure. Measuring sample size About 225 subjects were registered. This sample is considered large enough to estimate the incidence of relatively common adverse events with sufficient accuracy. Interim analysis Forms a DMC and is responsible for performing periodic reviews of cumulative safety data during the study period. DMC will operate independently of the sponsor and Quintiles and will be comprised of clinicians and biostatisticians with appropriate therapeutic expertise. DMC reviewed cumulative safety data for the tenepase catheter removal protocol at scheduled intervals, including studies of functionally abnormal CVA and HD catheters (study N3698g, N3 699g, N3700g, and N3 701g) and based on the results of the review process Responsible for making recommendations to the sponsor regarding the continued safety of the research. The specific guidelines and operational procedures of DMC will be summarized in the DMC regulations. Data Quality Assurance 126382.doc -108- 200826981

Quintiles provides a case report form (CRF) for this study. QuintUes is responsible for the data management of this trial, including dual data registration and quality inspection of the data. In the case of contradictory information, Quintiles will request resolution of site clarification. Quintiles will generate a Data Quality Plan that describes the quality checks to be performed on the CRF data. The CRF and calibration files will be indexed and imaged. Recording of system backups and research data stored in Quintiles will be in accordance with Quintiles' standard procedures. Use the QuintiieSi standard program experiment to send data directly to Quintiles to process and process the electronic transmission of such data.

Genentech will oversee the data management of the trial, including Quintnes data management and data quality design approval. The information will be transmitted electronically from Quintiles to Genentech on a regular basis and processed and processed electronically using Genentech's standard procedures. Safety Assessment The safety assessment will consist of monitoring and recording adverse events (AEs) and serious adverse events (SAEs) including target AEs. Adverse Events Regardless of attribution, AE is any unfavorable and unpleasant, addictive, symptomatic, or ill-related illness associated with the interventions applied to the product or the military program. It includes the following: • AEs that are not observed by previous persons during the AE reporting period specified in the protocol, complication, complications due to the intervention required by the protocol (eg, invasion such as biopsy) Presence & 乂 126382.doc 200826981 • Pre-existing medical condition (except for the condition being studied) that is worsened or qualitatively altered by the investigator as determined by the investigator during the AE reporting period specified in the protocol Severe adverse events If AE meets the following criteria, it is classified as SAE: • It causes death (ie, AE actually causes or causes death). • It is life-threatening (ie, the investigator believes that the subject is at risk of direct death. It does not include an AE that can lead to death if it exists in a more serious form). • It requires or extends hospitalization for inpatients. • It results in persistent or apparent disability/incapability (ie, the ability of the AE to cause the subject to perform normal life functions is generally disrupted). • It causes congenital anomalies/congenital defects in newborns/babies born to mothers exposed to the study drug. • It is a significant medical event considered by the investigator based on medical diagnosis (for example, a subject may be compromised or may require medical/surgical intervention to prevent one of the above results). An AE that does not meet any of these serious criteria should be considered a non-serious AE. 0 The terms "heavy" (&quot;severe") and "severe" ("seri〇us,") are not synonymous. Severity (or intensity) refers to a specific eight-rank rating, such as a mild "grade 丨" Moderate (grade 2) or severe (grade 3) myocardial infarction. ,, severely, for management = meaning (see previously defined) and based on the subject or event outcome or criteria of action that is usually associated with an event that poses a threat to the life or function of the subject. Strict 126382.doc -110- 200826981 Severity (non-severity) serves as a guide from the sponsor to the appropriate authority to define management reporting responsibilities. Severity and severity should be assessed independently when the person and SAE are recorded on the CRF. Target adverse event

Events that specifically cause specific attention (target AE) and include the following: • ICH major bleeding as evidenced by electrical tomography or magnetic resonance imaging, which is severely blood loss (&gt;5 mL/kg), requiring blood transfusion Blood loss or blood loss caused by low ash • Embolism, defined as any “severe” embolic event, including pulmonary events, arterial events (such as stroke, peripheral embolism or major organ embolism) or cholesterol plaques • thrombosis] including catheters Related venous thrombosis, defined as blood that causes pain, swelling, and/or ischemia of the extremities identified by radioimaging (eg, ultrasound, angiogram, or magnetic resonance) in the upper or lower extremity arteries or veins^ TABLE • CRBSI, further classified as follows: Clear μ: no other significant source of infection. Semi-quantitative cultures from the tip of the catheter (each catheter fragment > 15 colony forming units) and the same from peripheral or catheter blood samples The organism can be % type. No other obvious source of symptoms confirms the infection but the catheter tip has not confirmed the infection (or the catheter tip confirms that:: the liquid culture is not In the case of the case, with or without removal of the catheter, the symptom of fever after antibiotic treatment is 126382.doc -111 - 200826981 Potential type: no blood flow in the symptomatic subjects without other sources of eclipse The complication associated with symptomatic antipyretic after antibiotic treatment or removal of the catheter, as defined by the laboratory of the infection, is defined as rupture of the catheter during infusion or instillation of the drug, perforation of the indwelling vein, or surgical intervention (eg Hemorrhage at the catheter insertion site of suture or gauze wrap. The right ‘target AE satisfies the criteria outlined in Section 5丄2, which should be classified as SAE and should be reported as described in Section 5 4 . Methodology and timing for assessing and documenting safety variables The investigator is responsible for ensuring that all eight ugliness and SAE observed or reported during the study period. Adverse Event Reporting Cycle All AEs and SAEs must record a study period that begins at the start of the study treatment and ends after the second visit after the completion of the final study treatment or when the subject stops the study (any first) . Assessment of Adverse Events In addition to the follow-up visit on the 30th day, the investigator assessed the AEA SAE at each subject-assessment time point during the study period. All AEs and SAEs discovered by the investigator or detected by physical examination, laboratory tests or other methods during the inquiry will be recorded in the medical record of the subject and at the appropriate AE or On the SAE CRF page. The AE or SAE for each record will be governed by the duration (ie, start and end), severity (see Table 1), (if appropriate) severity criteria, and suspected relationship with research interests (see rules below) And take measures to describe. 126382.doc -112- 200826981 Table 1 Adverse event level (severity) scale severity describes mild temporary or mild discomfort (&lt;48 hours); does not interfere with the subject's regular activities; No need for medical intervention/therapy moderate to moderate to moderate interference with the subject's regular activities; no or minimal medical intervention/therapy required to cause considerable disruption to the subject's daily activities; medical intervention/therapy required; possible hospitalization Note: Regardless of severity, certain events may also meet management severity criteria. See the SAE definition in Section 5.1.2. To ensure consistency in the assessment of AE and SAE causes, the investigator will use the following general guidelines: • Yes There is a plausible temporary relationship between the AE episode and the study drug, and the AE cannot be easily affected by the subject's clinical condition, intervening disease, or accompanying The method states; and/or the AE meets the known pattern of response to the study drug; and/or the AE is relieved or eliminated after stopping the study drug or dose reduction, and (if appropriate) re-excited to reproduce. • The following evidence is absent: The AE has a source other than the study drug (eg, a pre-existing medical condition, a primary disease, an intermediate disease, or a concomitant drug); and/or the AE does not have a plausible temporary relationship with the administration of the study drug (eg, in the first One year after the study drug was given, the cancer was diagnosed.) 126382.doc -113 - 200826981 1 5 part of the document process. With the individual AE report, it is 丨丨 or,, non, f, such as the cause of the door The evaluation is irrelevant, and the organizer will immediately review the accumulated product experience.

All of the reported SAEs are confirmed and promptly communicated to the investigator and appropriate regulatory agencies. This new security raises adverse events.

All subjects should be assessed for a consistent method of time. Examples of non-directional queries include the following: How does π feel since the last clinical visit? , Has there been any new or changed since the last visit? &quot; Specific instructions for recording adverse events on the CRF § When recording ΑΕ or S ΑΕ on the CRF, research is poor and you should use the appropriate medical terminology/concept. Avoid proverbs and abbreviations. All ΑΕ should be recorded on the ^AE CRF page. A specified * cell exists on this page to indicate if the event is severe (γ/Ν). For sae, you must also complete the (4) CRF page. Only one medical concept should be recorded in the event area on the AE and SAE CRF pages. a• Relative to the diagnosis of symptoms and symptoms, right at the time of reporting, 6 know that diagnosis, but not individual symptoms and symptoms should be recorded on the ^ (for example, only &quot; liver failure or hepatitis instead of jaundice, flapping-like shock And transaminase upgrade). However, if the cluster of symptoms and/or symptoms is reported as a single diagnosis or symptom at the time of the report, the individual events should be recorded on the AE CRF page at 126382.doc -114- 200826981. If a diagnosis is subsequently determined, it should report the tracking information. b. Adverse events that occur after other events—generally, should be identified by the primary cause following other events (such as cascading events or clinical continuations). For example, if severe thirst is known to cause dehydration, it is sufficient to record only diarrhea on the AE CRF page. However, if a medically significant secondary AE is separated from the initiation event in time, both should be recorded as independent events. For example, if severe gastrointestinal bleeding causes kidney failure, then two events should be recorded on the separate AE CRF page. e•Continuous or recurrent adverse events The performance AE is an AE that continues to prolong between the time points of the assessment of the subject and cannot be eliminated. Unless the severity increases, such events should only be recorded in c叩: owed. If the persistence AE becomes more stringent, it should be recorded again on the Μ 页面 page. Recurrent AEs are AEs that appear, eliminate, and subsequently relapse. All recurrent AEs should be recorded on the AE CRF page. d. Clinical laboratory abnormalities Individual laboratory abnormalities were not recorded as AEs on CRF. Clinically significant laboratory abnormalities that only lead to study withdrawal, meet severity criteria, are related to clinical conditions or symptoms, or require medical intervention (eg, low hemoglobin requiring blood transfusion) are recorded on the AE CRF page. If a clinically significant laboratory abnormality is a symptom of a disease or syndrome (eg, alkaline phosphatase and bilirubin 5 times the upper limit of normal for cholecystitis), then only a diagnosis (eg, cholecystitis) should be recorded on the Ae CRF page. on. 126382.doc -115- 200826981 If a clinically significant laboratory abnormality is not a symptom of a disease or syndrome, the abnormality itself should be recorded on the AE CRF page. If a laboratory abnormality can be communicated as a clinical diagnosis, the diagnosis should be recorded as eight or sae. For example, a high serum potassium content of 7 〇 mEq/L should be recorded as, hyperkalemia,. Observations of the same clinically significant laboratory differences in each visit should not be repeated on the AE CRF page unless its severity, severity, or etiology changes. e. Pre-existing medical conditions Pre-existing medical conditions (IV) Medical conditions present at the beginning of the study. These conditions should be recorded on the cRF page of medical and surgical history. ^ Pre-existing medicines/conditions should be reassessed throughout the trial and recorded as sputum or SAE only if the frequency, severity, or characteristics of the disease deteriorated during the study. When recording such events on the AE CRF page, it is important to include the applicable descriptors to convey the concept that the pre-existing condition has changed such as 'a more frequent headache'.) f. Death has no attribution, scheme designation All deaths that occur during the AE reporting period will be recorded on the AE CRF page and promptly reported to the sponsor. When the death is recorded, the event or condition that caused or contributed to the fatal outcome should be recorded as a single medical concept on the AE CRF page. If the cause of death is unknown and cannot be confirmed at the time of the report, record "unclear death" on the AE CRF page. This situation should then be attempted to immediately identify the cause of death (eg, via a primary care physician, autopsy report, hospital record) and quickly report the cause of death to the sponsor. 126382.doc -116· 200826981 g. Any AE for hospitalization of medical or surgical procedures and a report of hospitalization or prolonged hospitalization for SAE. 2. The AE subject is hospitalized to undergo a medical or surgical procedure = the event of the procedure (4) The sequence itself should be recorded _. For example, if 2 = hospital to undergo coronary artery bypass surgery, the bypass will become a must-have heart disease recorded as SAE 〇

The following hospitalization materials were recorded on the CRF as:

And 2 records in the medical and surgical history of CRF II I &lt;

Hospitalization or prolonged residence of a b-surgery or elective surgical procedure p Treatment A Allows for efficacy of research (IV) Hospitalization or extended hospitalization of a predetermined therapy for a target disease of a hospitalization study in hospitalization 0 h·Pregnant If the female subject is pregnant within 9 days after receiving the study drug or the last dose of the study drug, the pregnancy report CRF should be completed and submitted promptly to the sponsor to facilitate the follow-up of the results. Sexual or spontaneous abortions should always be classified as serious, recorded as SAE and promptly reported to the sponsor. Similarly, any congenital anomalies/congenital defects of children born to female subjects exposed to the study drug should be recorded and Reported as SAE. i• If the post-study adverse event is due to prior study drug exposure, the investigator should notify the Medical Monitor to complete or stop the study by telephone 126382.doc •117-200826981 Any S AE that appears after participation. If the investigator finds out that the female subject who participated in the study subsequently gave birth to a cancer, or Abnormal, then who should notify the medical monitoring.

126382.doc 118- 200826981 Sequence Listing &lt;11〇&gt;US-based Nandek Company &lt;120&gt;Use of tissue plasminogen activator variants

&lt;130&gt; 39766-0242.TW &lt;140>096142107 &lt;141&gt; 2007-11-07 &lt;150&gt; 60/983,489 &lt;151&gt; 2007-10-29 &lt;150&gt; 60/864,758 &lt;151&gt; 2006-11-07 &lt;160&gt;2 &lt;170&gt; FastSEQ for Windows Version 4.0 &lt;210&gt; 1 &lt;211&gt;4 &lt;212&gt; PRT &lt;213&gt; Artificial Sequence &lt;220&gt;&lt;223&gt; Artificial Sequence &lt;400&gt; 1

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&lt;210 2 &lt;211&gt;4 &lt;212&gt; PRT &lt;213&gt;Artificial sequence &lt;220&gt;&lt;223&gt;Artificial sequence &lt;400>2 Ala Ala Ala Ala 1 126382.doc

Claims (1)

200826981 X. Patent Application Range: 1. A kit comprising a container comprising a solution comprising tenecteplase and the use of the solution to restore the function of a functional abnormal hemodialysis catheter present in a mammal. In the instructions, the catheter has a blood flow rate (BFR) of less than 300 ml/min, and the instructions "guide the user to locally deliver a total dose of about 2 to 4 mg of tenepase to all catheter lumens and allow Tenecteplase is left in the catheter for about 1 hour to about 72 hours so that the BFR of the catheter is no longer blocked. 2. The kit of claim 1 wherein during the 3 minute period prior to the hemodialysis The dysfunctional hemodialysis catheter with an arterial pressure of -250 mmHg additionally has a BFR of at least 25 mg/ml lower than the prescribed BFR. 3. The kit of claim 1 wherein the total dose of tenecteplase is about 4 mg. 4. The kit of any of claims 1 to 3, wherein the mammal is a human. 5. A method comprising the manufacture of a function for restoring a functionally abnormal hemodialysis catheter left in a mammal Naproxase, the catheter has a blood flow rate of less than 300 ml/min (^1^). 6. The method of claim 5, wherein the arterial pressure is -250 mmHg during 3 minutes before the hemodialysis The dysfunctional hemodialysis catheter additionally has a BFR that is at least 25 mg/ml lower than the prescribed BFR. 7. The method of claim 5, further comprising direct administration of a total dose of about 2 to 4 mg of tenepase. Into all catheter lumens, and allowed to remain in the catheter for about 1 hour to about 72 hours so that the BFR of the catheter is no longer obstructed. 126382.doc 200826981 8. 9. 10. 11. 12. 13. 14. The method of any one of claims 5 to 7, wherein the mammal is a human. A tenecteplase enzyme for restoring dysfunctional blood left in a mammalian order The function of a dialysis catheter having a blood flow rate (BFR) of less than 3 ml/min. For example, the tenecteplase of claim 9 wherein during the period of the hemodialysis, the arterial pressure is -250 mmHg. This functional abnormal hemodialysis guide has an additional lower than the prescription Bfr by at least 25 For example, the tenectease of claim 9, wherein the tenecteplase is in a solution for sterilizing water for injection or bacteriostatic water for injection, such as tenectease according to item 9, wherein Tenepase is retained in the catheter until the BFR of the catheter is increased beyond the BFR prior to administration of tenecteplase and the increase is maintained for at least 48 hours. As described in claim 9, the tenecteplase, wherein The nepase is sterilized in water. Θ The tenecteplase of claim 9, wherein the total dose of tenecteplase is about 4 mg. The tenecteplase of claim 9, wherein the tenecteplase is dropped into the catheter for about one hour. For example, the tenecteplase of claim 9, wherein the tenecteplase is instilled into the catheter in an extended indwelling form of from 1 hour to about 72 hours. : The tenecteplase of claim 16, wherein the indwelling is from about 2 to about as small as the tenth of tenth of tenectilase in which the tenectilase is administered - wherein the tenecteplase is administered - 126382.doc 200826981 19. The tenectease of claim 18 is re-experienced; the middle line is administered to the mammal during the dialysis treatment period. The enzyme is administered by one or 20. The tenectease of claim 9, wherein the tenectin is a tenectease according to the formula 2, wherein the tenecteplase is administered once. 22. The tenecteplase of claim 9, wherein the mammal undergoes hemodialysis after administration of the tenecteplase. The tenecteplase according to any one of claims 9 to 22, wherein the mammal is a human. 126382.doc 200826981 VII. Designation of Representative Representatives (1) The representative representative of the case is: (none) (2) The symbol of the symbol of the representative figure is simple: f VIII. If there is a chemical formula in this case, please reveal the best display invention. Characteristic chemical formula: (none) 126382.doc